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Sample records for superficial actinic porokeratosis

  1. Localised nasal actinic porokeratosis: An independent subtype of porokeratosis?

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    Nidhi Sharma

    2017-01-01

    Full Text Available Porokeratosis (PK are a group of keratinizing disorders clinically characterized by atrophic plaques with peripheral hyperkeratotic ridge which gradually spread in a centrifugal manner. The sites of predilection are trunk or extremities and facial lesions are rarely reported. We report a sporadic adult-onset case of porokeratosis of a 20 year old female with a solitary, asymptomatic, persistent classical plaque confined to the ala of nose. On reviewing the literature, nasal and perinasal area has been found to be exclusively involved in many cases of facial porokeratosis. Such cases have unique yet common epidemiological and clinical profile thereby deserving a separate category while being classified.

  2. A Case of Porokeratosis Showing Different Clinical Patterns of the Disease with Anogenital Involvement

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    Özlem Karabudak

    2008-10-01

    Full Text Available Porokeratosis (PK is a group of cutaneous entities characterized by marginate scaling lesions, histologically showing a column of parakeratotic keratinocytes (cornoid lamella. Various forms are recognized such as porokeratosis of Mibelli (PM, linear porokeratosis, disseminated superficial actinic porokeratosis, punctate parakeratosis. PM should be treated because of the possibility of developing malignant epithelial tumors. We are presenting a 21 year old male patient suffering from PM on the back of the hands, foot, scrotum, oral mucosa and anal region. The histological biopsy specimens showed the characteristic features of porokeratosis. We destroyed the lesions by cryotherapy sessions. Here, we present a case of PM since it is rarely seen as multiple lesions with oral, anal and scrotal involvements altogether. (Turkderm 2008; 42: 97-9

  3. Disease: H00791 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available H00791 Disseminated superficial actinic porokeratosis (DSAP) Porokeratosis is a di...amella. Disseminated superficial actinic porokeratosis (DSAP) is the most common form of porokeratosis. It i... (description) ... AUTHORS ... Sertznig P, von Felbert V, Megahed M ... TITLE ... Porokeratosis...isseminated superficial actinic porokeratosis in a patient undergoing treatment w...eminated superficial and warty type of porokeratosis: a rare coexistence. ... JOURNAL ... Indian J Dermatol 56:576-7 (2011) DOI:10.4103/0019-5154.87160 ...

  4. Hyperkeratotic porokeratosis ptychotropica with satellite lesions: a rare presentation of an unusual variant of porokeratosis.

    Science.gov (United States)

    Hazarika, Debeeka; Pawar, Manoj

    2017-12-01

    Since its description in 1995, porokeratosis ptychotropica (PP) has remained a less-recognized variant of porokeratosis (PK). The term porokeratosis ptychotropica was coined in reference to its characteristic of affecting body folds. It mimics many other dermatological diseases and is therefore often misdiagnosed. We report a patient with multiple hyperkeratotic, warty lesions across the buttocks that mimicked cutaneous tuberculosis (CTB), but histological examination confirmed the correct diagnosis of PP.

  5. Patient Perception of Imiquimod Treatment for Actinic Keratosis and Superficial Basal Cell Carcinoma in 202 Patients

    NARCIS (Netherlands)

    Waalboer-Spuij, Rick; Holterhues, Cynthia; van Hattem, Simone; Schuttelaar, Marie Louise A.; Gaastra, Menno T. W.; Kuijpers, Danielle I. M.; Hollestein, Loes M.; Nijsten, Tamar E. C.

    2015-01-01

    Objective: To document the impact on patient-reported outcomes and health-related quality of life (HRQoL) of treatment with imiquimod cream in patients with actinic keratosis (AK) and superficial basal cell carcinoma (sBCC). Methods: This open-label, multicenter study included AK and sBCC patients

  6. Dermal changes in superficial basal cell carcinoma, melanoma in situ and actinic keratosis and their implications

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    Kazlouskaya, Viktoryia; Malhotra, Saurabh; Navarro, Raquel; Wu, Karen Nguyen; Shvartsbeyn, Marianna; Shengli, Chen; Gui, Jiang; Elston, Dirk M.

    2018-01-01

    Background Basal cell carcinoma (BCC) has a characteristic stroma, but less is known about the dermal characteristics associated with melanoma in situ (MIS) and actinic keratosis (AK). Materials and methods Dermal changes were studied in 301 specimens of AK, BCC and MIS. Subsequently, blinded images of dermal changes from 90 randomly selected cases of those entities were used to assess the predictive value of the dermal changes. Agreement with the final diagnosis was calculated using kappa coefficient (κ). Results Fibromyxoid stroma was present in 82% of BCC cases; fibrous stroma was seen in 25% of BCC, 58% of MIS and 35.6% of AK specimens (p <0.05). A lichenoid inflammatory infiltrate was frequently associated with AK and a perifollicular infiltrate with periadnexal fibrosis with MIS. Blinded evaluation of images of the dermal changes associated with the tumors yielded the correct diagnosis in (54.4, 41.1 and 27.8%; average 41.2%) by the three appraisers. Coefficient of agreement in blinded imaged evaluation with the actual diagnosis was higher in the BCC and MIS compared with AK (κ = 0.37, p = 0.0001; κ = 0.2, p = 0.0005 and κ = −0.06, p = 0.84, respectively). Conclusion Dermal features may be helpful in predicting the correct diagnosis when tumor is not visible. PMID:24117926

  7. Familial disseminated plaque type porokeratosis with multiple horns and squamous cell carcinoma involving anal skin

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    Sarma Nilendu

    2009-01-01

    Full Text Available Porokeratosis is a disorder of keratinization showing a well-defined lesion with a hyperkeratotic ridge on the border that contains the coronoid lamella. We report familial (autosomal dominant with reduced penetrance disseminated plaque type (Mibelli′s type porokeratosis in a father and son. In the father, there were multiple horns and a large squamous cell carcinoma in a large lesion over the perianal region that reached up to the squamo-columnar junction of the anal mucosa and even invaded the anal sphincteric muscles. Disseminated lesions of the Mibelli′s type, development of horns, and malignancy in this unusual location have not been previously reported.

  8. Actinic keratosis

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    Solar keratosis; Sun-induced skin changes - keratosis; Keratosis - actinic (solar); Skin lesion - actinic keratosis ... Actinic keratosis is caused by exposure to sunlight. You are more likely to develop it if you: Have fair ...

  9. Wong-Type Dermatomyositis Showing Porokeratosis-Like Changes (Columnar Dyskeratosis: A Case Report and Review of the Literature

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    Nicole Umanoff

    2015-01-01

    Full Text Available Background: Wong-type dermatomyositis (DM exhibits simultaneous pityriasis rubra pilaris (PRP features. Case Report: A 50-year-old woman presented with a heliotrope rash, Gottron's papules, and a poikilodermic, erythematous rash in shawl distribution without evidence of muscle weakness. Despite topical corticosteroids, the eruption progressed 9 months later to include generalized hyperkeratotic follicular papules, islands of sparing, and atrophic macules with a collarette of scale suggestive of porokeratosis. Mild dysphonia was the only sign of muscle weakness. Serology showed positive ANA. Histopathology revealed interface dermatitis with dermal mucin and melanophages, irregular psoriasiform hyperplasia, alternating mounds of para- and orthokeratosis, and tiers of dyskeratotic cells (columnar dyskeratosis. Systemic corticosteroid therapy was not tolerated; acitretin diminished the hyperkeratosis. While hyperpigmentation persisted, no progression of cutaneous or muscular symptoms has occurred after 22 months of follow-up and cessation of the therapy. Overall, her course did not differ from the natural history documented in the literature review of Wong-type DM. The most similar case also exhibited pseudocornoid lamella changes. Conclusion: Wong-type DM is a clinicopathologic DM-PRP hybrid that can also exhibit porokeratosis-like features best described as columnar dyskeratosis. Recognizing these types of lesions in DM is warranted in order to make an accurate assessment of their prognostic significance.

  10. Actinic keratosis

    DEFF Research Database (Denmark)

    Erlendsson, Andrés M; Egekvist, Henrik; Lorentzen, Henrik F.

    2016-01-01

    Objectives: The incidence of actinic keratosis (AK) is increasing, and several treatment options are available. The aim of this study was to describe clinical characteristics and treatment patterns in patients with AK treated by Danish dermatologists. Methods: A multicenter, non-interventional, c......Objectives: The incidence of actinic keratosis (AK) is increasing, and several treatment options are available. The aim of this study was to describe clinical characteristics and treatment patterns in patients with AK treated by Danish dermatologists. Methods: A multicenter, non...... and currently suspected in 9.4% of AK-affected anatomical regions. Lesions of AK were located primarily on the face (38.6%), scalp (12.8%), and hands (11.2%). Actinic keratosis commonly presented with multiple AK lesions (38.6%) and field cancerization (38.5%). The treatments used most frequently were...

  11. Bacterial Actins.

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    Izoré, Thierry; van den Ent, Fusinita

    2017-01-01

    A diverse set of protein polymers, structurally related to actin filaments contributes to the organization of bacterial cells as cytomotive or cytoskeletal filaments. This chapter describes actin homologs encoded by bacterial chromosomes. MamK filaments, unique to magnetotactic bacteria, help establishing magnetic biological compasses by interacting with magnetosomes. Magnetosomes are intracellular membrane invaginations containing biomineralized crystals of iron oxide that are positioned by MamK along the long-axis of the cell. FtsA is widespread across bacteria and it is one of the earliest components of the divisome to arrive at midcell, where it anchors the cell division machinery to the membrane. FtsA binds directly to FtsZ filaments and to the membrane through its C-terminus. FtsA shows altered domain architecture when compared to the canonical actin fold. FtsA's subdomain 1C replaces subdomain 1B of other members of the actin family and is located on the opposite side of the molecule. Nevertheless, when FtsA assembles into protofilaments, the protofilament structure is preserved, as subdomain 1C replaces subdomain IB of the following subunit in a canonical actin filament. MreB has an essential role in shape-maintenance of most rod-shaped bacteria. Unusually, MreB filaments assemble from two protofilaments in a flat and antiparallel arrangement. This non-polar architecture implies that both MreB filament ends are structurally identical. MreB filaments bind directly to membranes where they interact with both cytosolic and membrane proteins, thereby forming a key component of the elongasome. MreB filaments in cells are short and dynamic, moving around the long axis of rod-shaped cells, sensing curvature of the membrane and being implicated in peptidoglycan synthesis.

  12. Actin genes and their expression in pacific white shrimp, Litopenaeus vannamei.

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    Zhang, Xiaoxi; Zhang, Xiaojun; Yuan, Jianbo; Du, Jiangli; Li, Fuhua; Xiang, Jianhai

    2018-04-01

    Actin is a multi-functional gene family that can be divided into muscle-type actins and non-muscle-type actins. In this study, 37 unigenes encoding actins were identified from RNA-Seq data of Pacific white shrimp, Litopenaeus vannamei. According to phylogenetic analysis, four and three cDNAs belong to cytoplasmic- and heart-type actins and were named LvActinCT and LvActinHT, respectively. 10 cDNAs belong to the slow-type skeletal muscle actins, and 18 belong to the fast-type skeletal muscle actins; they were designated LvActinSSK and LvActinFSK, respectively. Some muscle actin genes formed gene clusters in the genome. Multiple alternative transcription starts sites (ATSSs) were found for LvActinCT1. Based on the early developmental expression profile, almost all LvActins were highly expressed between the early limb bud and post-larval stages. Using LvActinSSK5 as probes, slow-type muscle was localized in pleopod muscle and superficial ventral muscle. We also found three actin genes that were down-regulated in the hemocytes of white spot syndrome virus (WSSV)- and Vibrio parahaemolyticus-infected L. vannamei. This study provides valuable information on the actin gene structure of shrimp, furthers our understanding of the shrimp muscle system and helps us develop strategies for disease control and sustainable shrimp farming.

  13. Actinic Keratosis Pathogenesis Update and New Patents.

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    Cantisani, Carmen; Paolino, Giovanni; Melis, Marcello; Faina, Valentina; Romaniello, Federico; Didona, Dario; Cardone, Michele; Calvieri, Stefano

    2016-01-01

    Actinic keratosis is a common premalignant skin lesion. Because of its increasing incidence, several efforts have been made to earlier detectection and to improve knowledge on photocarcinogenic pathways of keratinocytes. As a consequence, recently new discoveries have been done in this field. Starting from our previous review on actinic keratosis, we reviewed the literature focusing on pathogenesis and new patents in order to highlight the most recent progresses in diagnosis and therapeutic approach. Although several efforts have been done in the field of photodamaged skin, new upgrades in diagnosis and therapy are needed to detect superficial actinic keratosis earlier, to improve the disease free survival of patient and to better treat the field cancerization.

  14. Diclofenac Topical (actinic keratosis)

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    ... topical gel (Solaraze) is used to treat actinic keratosis (flat, scaly growths on the skin caused by ... The way diclofenac gel works to treat actinic keratosis is not known.Diclofenac is also available as ...

  15. Superficial skin ulcers

    International Nuclear Information System (INIS)

    Samaila, Modupeola O.; Rafindadi, Abdulmumini H.; Oluwole, Olabode P.; Adewuyi, Sunday A.

    2007-01-01

    Objective was to determine the underlying cause of superficial skin ulcers over a 15-year period. A retrospective histopathological analysis of 670 cases of superficial skin ulcers diagnosed in the Dept. of Pathology, Ahmadu Bello University Teaching Hospital, Zaria, Nigeria from January 1991 to December 2005. A total of 670 superficial skin ulcers were analyzed. The mail to female gender ratio was 409:261(1.5:1.0) and a peakage frequency of 44.3 %( 297) in the 5th and 6th decades. Spectrum of lesions encountered was categorized into inflammatory, infections, benign and malignant diseases. The malignant lesions were 309 (46.1%), non-specific inflammation 302 (45.1%), granulation tissue 25 (3.7%) and pseudoepitheliomatous hyperplasia 14 (2.1%). A total of 18(2.7%) specific infections were encountered, which included bacterial, fungal and viral infection. Benign lesions were 2(0.3%), comprising of neurofibroma and Bowen's disease. The most common malignant lesion was squamous cell carcinoma 203 (30.3%) with a male to female ratio of 128:75 (1.7:1.0). Of these 161 were well differentiated tumors. The lower limb was the prevalent site distribution of all the ulcers. Superficial ulcers may be harbinger of malignant diseases. Squamous cell carcinoma remains the most common malignant lesion arising from chronic superficial ulcers from our setting. Adequate tissue biopsy and early diagnosis may reduce the attendant morbidity of these ulcers. (author)

  16. Polycation induced actin bundles

    OpenAIRE

    Muhlrad, Andras; Grintsevich, Elena E.; Reisler, Emil

    2011-01-01

    Three polycations, polylysine, the polyamine spermine and the polycationic protein lysozyme were used to study the formation, structure, ionic strength sensitivity and dissociation of polycation-induced actin bundles. Bundles form fast, simultaneously with the polymerization of MgATP-G-actins, upon addition of polycations to solutions of actins at low ionic strength conditions. This indicates that nuclei and/or nascent filaments bundle due to attractive, electrostatic effect of polycations an...

  17. Superficies de segundo orden

    OpenAIRE

    Salazar Salazar, Luis Álvaro

    1987-01-01

    Este trabajo se propone poner al alcance de estudiantes de primeros semestres de carreras de aplicación de la matemática, un algoritmo proporcionado por el álgebra lineal, para tratar con mas generalidad, agilidad y libertad unos objetos de la geometría analítica de no fácil manipulación por otros métodos y que se conocen como superficies de segundo orden o superficies cuádricas. En este orden de ideas, el autor considera importante que con este tratamiento se incluya este tema en una asignat...

  18. Topical Imiquimod in the Treatment of Conjunctival Actinic Keratosis.

    Science.gov (United States)

    Rowlands, Megan A; Giacometti, Joseph N; Servat, Javier; Materin, Miguel A; Levin, Flora

    Conjunctival actinic keratosis is rare and difficult to treat, as recurrences are common. Imiquimod, an immune response modulator, is currently Food and Drug Administration-approved for cutaneous actinic keratosis and superficial basal cell carcinomas. Emerging reports have shown it to be effective in treating some periocular and conjunctival lesions. The authors present a case of a 68-year-old white man with recurrent actinic keratosis involving the pretarsal conjunctiva, which was successfully treated with 5% topical imiquimod following previous failure with cryotherapy and interferon α-2b. The patient had ocular irritation that resolved on cessation of treatment. To the authors' knowledge, this is the first report of conjunctival actinic keratosis being treated with and successfully eradicated by topical imiquimod.

  19. Common Superficial Bursitis.

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    Khodaee, Morteza

    2017-02-15

    Superficial bursitis most often occurs in the olecranon and prepatellar bursae. Less common locations are the superficial infrapatellar and subcutaneous (superficial) calcaneal bursae. Chronic microtrauma (e.g., kneeling on the prepatellar bursa) is the most common cause of superficial bursitis. Other causes include acute trauma/hemorrhage, inflammatory disorders such as gout or rheumatoid arthritis, and infection (septic bursitis). Diagnosis is usually based on clinical presentation, with a particular focus on signs of septic bursitis. Ultrasonography can help distinguish bursitis from cellulitis. Blood testing (white blood cell count, inflammatory markers) and magnetic resonance imaging can help distinguish infectious from noninfectious causes. If infection is suspected, bursal aspiration should be performed and fluid examined using Gram stain, crystal analysis, glucose measurement, blood cell count, and culture. Management depends on the type of bursitis. Acute traumatic/hemorrhagic bursitis is treated conservatively with ice, elevation, rest, and analgesics; aspiration may shorten the duration of symptoms. Chronic microtraumatic bursitis should be treated conservatively, and the underlying cause addressed. Bursal aspiration of microtraumatic bursitis is generally not recommended because of the risk of iatrogenic septic bursitis. Although intrabursal corticosteroid injections are sometimes used to treat microtraumatic bursitis, high-quality evidence demonstrating any benefit is unavailable. Chronic inflammatory bursitis (e.g., gout, rheumatoid arthritis) is treated by addressing the underlying condition, and intrabursal corticosteroid injections are often used. For septic bursitis, antibiotics effective against Staphylococcus aureus are generally the initial treatment, with surgery reserved for bursitis not responsive to antibiotics or for recurrent cases. Outpatient antibiotics may be considered in those who are not acutely ill; patients who are acutely ill

  20. SUPERFICIAL CERVICAL PLEXUS BLOCK

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    Komang Mega Puspadisari

    2014-01-01

    Full Text Available Superficial cervical plexus block is one of the regional anesthesia in  neck were limited to thesuperficial fascia. Anesthesia is used to relieve pain caused either during or after the surgery iscompleted. This technique can be done by landmark or with ultrasound guiding. The midpointof posterior border of the Sternocleidomastoid was identified and the prosedure done on thatplace or on the level of cartilage cricoid.

  1. Histones bundle F-actin filaments and affect actin structure.

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    Edna Blotnick

    Full Text Available Histones are small polycationic proteins complexed with DNA located in the cell nucleus. Upon apoptosis they are secreted from the cells and react with extracellular polyanionic compounds. Actin which is a polyanionic protein, is also secreted from necrotic cells and interacts with histones. We showed that both histone mixture (histone type III and the recombinant H2A histone bundles F-actin, increases the viscosity of the F-actin containing solution and polymerizes G-actin. The histone-actin bundles are relatively insensitive to increase of ionic strength, unlike other polycation, histatin, lysozyme, spermine and LL-37 induced F-actin bundles. The histone-actin bundles dissociate completely only in the presence of 300-400 mM NaCl. DNA, which competes with F-actin for histones, disassembles histone induced actin bundles. DNase1, which depolymerizes F- to G-actin, actively unbundles the H2A histone induced but slightly affects the histone mixture induced actin bundles. Cofilin decreases the amount of F-actin sedimented by low speed centrifugation, increases light scattering and viscosity of F-actin-histone mixture containing solutions and forms star like superstructures by copolymerizing G-actin with H2A histone. The results indicate that histones are tightly attached to F-actin by strong electrostatic and hydrophobic forces. Since both histones and F-actin are present in the sputum of patients with cystic fibrosis, therefore, the formation of the stable histone-actin bundles can contribute to the pathology of this disease by increasing the viscosity of the sputum. The actin-histone interaction in the nucleus might affect gene expression.

  2. Histones bundle F-actin filaments and affect actin structure.

    Science.gov (United States)

    Blotnick, Edna; Sol, Asaf; Muhlrad, Andras

    2017-01-01

    Histones are small polycationic proteins complexed with DNA located in the cell nucleus. Upon apoptosis they are secreted from the cells and react with extracellular polyanionic compounds. Actin which is a polyanionic protein, is also secreted from necrotic cells and interacts with histones. We showed that both histone mixture (histone type III) and the recombinant H2A histone bundles F-actin, increases the viscosity of the F-actin containing solution and polymerizes G-actin. The histone-actin bundles are relatively insensitive to increase of ionic strength, unlike other polycation, histatin, lysozyme, spermine and LL-37 induced F-actin bundles. The histone-actin bundles dissociate completely only in the presence of 300-400 mM NaCl. DNA, which competes with F-actin for histones, disassembles histone induced actin bundles. DNase1, which depolymerizes F- to G-actin, actively unbundles the H2A histone induced but slightly affects the histone mixture induced actin bundles. Cofilin decreases the amount of F-actin sedimented by low speed centrifugation, increases light scattering and viscosity of F-actin-histone mixture containing solutions and forms star like superstructures by copolymerizing G-actin with H2A histone. The results indicate that histones are tightly attached to F-actin by strong electrostatic and hydrophobic forces. Since both histones and F-actin are present in the sputum of patients with cystic fibrosis, therefore, the formation of the stable histone-actin bundles can contribute to the pathology of this disease by increasing the viscosity of the sputum. The actin-histone interaction in the nucleus might affect gene expression.

  3. Actinic keratosis among seafarers.

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    Oldenburg, M; Kuechmeister, B; Ohnemus, U; Baur, X; Moll, I

    2013-11-01

    The aim of this study was to assess the prevalence of UV-induced actinic keratosis and further skin lesions. A newly developed questionnaire about lifetime UV radiation exposure was completed by 514 seafarers. An experienced dermatologist inspected the whole-body skin status of all participants. The questionnaire revealed a pre-employment UV radiation exposure in 104 seafarers, sunbed use in 26 subjects and a median work-related UV radiation exposure at sea of 20 years. The diagnosis of actinic keratoses was made in 94 seafarers and the clinical diagnosis of skin cancers in 48 seafarers (28 basal cell carcinoma, 11 squamous cell carcinoma, 9 malignant melanoma). After age standardisation according to a European reference population, the male European seafarers in this study had a 1.80-fold increased risk of actinic keratosis. Actinic keratoses [OR 1.03 (1.01-1.05)] and squamous cell carcinoma [OR 1.07 (1.01-1.13)] were related to the duration of seafaring time in years. A significant association was also found between actinic keratosis/squamous cell carcinoma and sunlight exposure during home leave [OR 1.67 (1.03-2.81) and OR 6.19 (1.18-32.40)]. Furthermore, the engine room personnel-especially the technical officers-were at higher risk of developing actinic keratosis. Due to the high prevalence of actinic keratosis especially among older seafarers with fair skin, with longer duration of seafaring employment at sea and with higher UV exposure during home leave, more intensive advice should be given on sun protection both at sea and ashore.

  4. Polycation induced actin bundles.

    Science.gov (United States)

    Muhlrad, Andras; Grintsevich, Elena E; Reisler, Emil

    2011-04-01

    Three polycations, polylysine, the polyamine spermine and the polycationic protein lysozyme were used to study the formation, structure, ionic strength sensitivity and dissociation of polycation-induced actin bundles. Bundles form fast, simultaneously with the polymerization of MgATP-G-actins, upon the addition of polycations to solutions of actins at low ionic strength conditions. This indicates that nuclei and/or nascent filaments bundle due to attractive, electrostatic effect of polycations and the neutralization of repulsive interactions of negative charges on actin. The attractive forces between the filaments are strong, as shown by the low (in nanomolar range) critical concentration of their bundling at low ionic strength. These bundles are sensitive to ionic strength and disassemble partially in 100 mM NaCl, but both the dissociation and ionic strength sensitivity can be countered by higher polycation concentrations. Cys374 residues of actin monomers residing on neighboring filaments in the bundles can be cross-linked by the short span (5.4Å) MTS-1 (1,1-methanedyl bismethanethiosulfonate) cross-linker, which indicates a tight packing of filaments in the bundles. The interfilament cross-links, which connect monomers located on oppositely oriented filaments, prevent disassembly of bundles at high ionic strength. Cofilin and the polysaccharide polyanion heparin disassemble lysozyme induced actin bundles more effectively than the polylysine-induced bundles. The actin-lysozyme bundles are pathologically significant as both proteins are found in the pulmonary airways of cystic fibrosis patients. Their bundles contribute to the formation of viscous mucus, which is the main cause of breathing difficulties and eventual death in this disorder. Copyright © 2011 Elsevier B.V. All rights reserved.

  5. Superficial fungal infections.

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    Schwartz, Robert A

    Superficial fungal infections arise from a pathogen that is restricted to the stratum corneum, with little or no tissue reaction. In this Seminar, three types of infection will be covered: tinea versicolor, piedra, and tinea nigra. Tinea versicolor is common worldwide and is caused by Malassezia spp, which are human saprophytes that sometimes switch from yeast to pathogenic mycelial form. Malassezia furfur, Malassezia globosa, and Malassezia sympodialis are most closely linked to tinea versicolor. White and black piedra are both common in tropical regions of the world; white piedra is also endemic in temperate climates. Black piedra is caused by Piedraia hortae; white piedra is due to pathogenic species of the Trichosporon genus. Tinea nigra is also common in tropical areas and has been confused with melanoma.

  6. Actin, actin-binding proteins, and actin-related proteins in the nucleus.

    Science.gov (United States)

    Kristó, Ildikó; Bajusz, Izabella; Bajusz, Csaba; Borkúti, Péter; Vilmos, Péter

    2016-04-01

    Extensive research in the past decade has significantly broadened our view about the role actin plays in the life of the cell and added novel aspects to actin research. One of these new aspects is the discovery of the existence of nuclear actin which became evident only recently. Nuclear activities including transcriptional activation in the case of all three RNA polymerases, editing and nuclear export of mRNAs, and chromatin remodeling all depend on actin. It also became clear that there is a fine-tuned equilibrium between cytoplasmic and nuclear actin pools and that this balance is ensured by an export-import system dedicated to actin. After over half a century of research on conventional actin and its organizing partners in the cytoplasm, it was also an unexpected finding that the nucleus contains more than 30 actin-binding proteins and new classes of actin-related proteins which are not able to form filaments but had evolved nuclear-specific functions. The actin-binding and actin-related proteins in the nucleus have been linked to RNA transcription and processing, nuclear transport, and chromatin remodeling. In this paper, we attempt to provide an overview of the wide range of information that is now available about actin, actin-binding, and actin-related proteins in the nucleus.

  7. Chronic Actinic Dermatitis

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    Bengü Çevirgen Cemil

    2017-06-01

    Full Text Available Chronic actinic dermatitis (CAD is characterized by persistent eczema-like lesions, mainly on sun-exposed sites, induced by ultraviolet B, sometimes ultraviolet A, and occasionally visible light. CAD is a rare photodermatitis. It is often associated with contact allergens including airborne allergens such as fragrances, plant antigens and topical medications. A 62 year old farmer is applied with eczematous lesions restricted to sun-exposed areas. Clinical findings and histopathologic features were consistent with the diagnosis of chronic actinic dermatitis. The patient also had contact allergy to multiple allergens. We present this case to emphasize the significance of patch test on CAD treatment and the success of topical tacrolimus and azathioprine.

  8. Actinic reticuloid. Diagnostics

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    E. V. Sokolovskiy

    2016-01-01

    Full Text Available This article is about the case of actinic reticuloid - the rare dermatosis which clinical presentation is similar to atopic dermatitis, T-cell lymphoma. Good treatment effect was obtained by long cycles (2 cycles for 3 months of hydroxychloroquine and sun protective therapy included sunscreens SPF 50, nicotinic acid, sun-safe clothes which blocked ultraviolet radiation without any glucocorticosteroid drugs and cytostatic treatment.

  9. Actin filaments as tension sensors.

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    Galkin, Vitold E; Orlova, Albina; Egelman, Edward H

    2012-02-07

    The field of mechanobiology has witnessed an explosive growth over the past several years as interest has greatly increased in understanding how mechanical forces are transduced by cells and how cells migrate, adhere and generate traction. Actin, a highly abundant and anomalously conserved protein, plays a large role in forming the dynamic cytoskeleton that is so essential for cell form, motility and mechanosensitivity. While the actin filament (F-actin) has been viewed as dynamic in terms of polymerization and depolymerization, new results suggest that F-actin itself may function as a highly dynamic tension sensor. This property may help explain the unusual conservation of actin's sequence, as well as shed further light on actin's essential role in structures from sarcomeres to stress fibers. Copyright © 2012 Elsevier Ltd. All rights reserved.

  10. The effects of near-UV radiation on elasmobranch lens cytoskeletal actin.

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    Zigman, S; Rafferty, N S; Scholz, D L; Lowe, K

    1992-08-01

    The role of near-UV radiation as a cytoskeletal actin-damaging agent was investigated. Two procedures were used to analyse fresh smooth dogfish (Mustelus canis) eye lenses that were incubated for up to 22 hr in vitro, with elasmobranch Ringer's medium, and with or without exposure to a near-UV lamp (emission principally at 365 nm; irradiance of 2.5 mW cm-2). These were observed histologically using phalloidin-rhodamine specific staining and by transmission electron microscopy. In addition, solutions of purified polymerized rabbit muscle actin were exposed to the same UV conditions and depolymerization was assayed by ultracentrifugation and high-pressure liquid chromatography. While the two actins studied do differ very slightly in some amino acid sequences, they would react physically nearly identically. The results showed that dogfish lenses developed superficial opacities due to near-UV exposure. Whole mounts of lens epithelium exhibited breakdown of actin filaments in the basal region of the cells within 18 hr of UV exposure. TEM confirmed the breakdown of actin filaments due to UV exposure. SDS-PAGE and immunoblotting positively identified actin in these cells. Direct exposure of purified polymerized muscle actin in polymerizing buffer led to an increase in actin monomer of approximately 25% in the UV-exposed solutions within 3-18 hr, whether assayed by ultracentrifugation or HPLC. The above indicates that elasmobranch lens epithelial cells contain UV-labile actin filaments, and that near-UV radiation, as is present in the sunlit environment, can break down the actin structure in these cells. Furthermore, breakdown of purified polymerized muscle actin does occur due to near-UV light exposure.(ABSTRACT TRUNCATED AT 250 WORDS)

  11. Superficial Priming in Episodic Recognition

    Science.gov (United States)

    Dopkins, Stephen; Sargent, Jesse; Ngo, Catherine T.

    2010-01-01

    We explored the effect of superficial priming in episodic recognition and found it to be different from the effect of semantic priming in episodic recognition. Participants made recognition judgments to pairs of items, with each pair consisting of a prime item and a test item. Correct positive responses to the test item were impeded if the prime…

  12. Superficial parotidectomy via facelift incision

    NARCIS (Netherlands)

    Lohuis, Peter J. F. M.; Tan, M. Liane; Bonte, Katrien; van den Brekel, Michiel W. M.; Balm, Alfons J. M.; Vermeersch, Hubert B.

    2009-01-01

    The stigma of a visually prominent facial scar following parotid surgery can be distressing to a young patient. The surgical technique of parotidectomy via a facelift incision is described and evaluated. Thirty patients with a benign lesion of the parotid gland underwent a partial superficial

  13. Interventions for actinic keratoses.

    Science.gov (United States)

    Gupta, Aditya K; Paquet, Maryse; Villanueva, Elmer; Brintnell, William

    2012-12-12

    Actinic keratoses are a skin disease caused by long-term sun exposure, and their lesions have the potential to develop into squamous cell carcinoma. Treatments for actinic keratoses are sought for cosmetic reasons, for the relief of associated symptoms, or for the prevention of skin cancer development. Detectable lesions are often associated with alteration of the surrounding skin (field) where subclinical lesions might be present. The interventions available for the treatment of actinic keratoses include individual lesion-based (e.g. cryotherapy) or field-directed (e.g. topical) treatments. These might vary in terms of efficacy, safety, and cosmetic outcomes. To assess the effects of topical, oral, mechanical, and chemical interventions for actinic keratosis. We searched the following databases up to March 2011: the Cochrane Skin Group Specialised Register, CENTRAL in The Cochrane Library, MEDLINE (from 2005), EMBASE (from 2010), and LILACS (from 1982). We also searched trials registers, conference proceedings, and grey literature sources. Randomised controlled trials (RCTs) comparing the treatment of actinic keratoses with either placebo, vehicle, or another active therapy. At least two authors independently abstracted data, which included adverse events, and assessed the quality of evidence. We performed meta-analysis to calculate a weighted treatment effect across trials, and we expressed the results as risk ratios (RR) and 95% confidence intervals (CI) for dichotomous outcomes (e.g. participant complete clearance rates), and mean difference (MD) and 95% CI for continuous outcomes (e.g. mean reduction in lesion counts). We included 83 RCTs in this review, with a total of 10,036 participants. The RCTs covered 18 topical treatments, 1 oral treatment, 2 mechanical interventions, and 3 chemical interventions, including photodynamic therapy (PDT). Most of the studies lacked descriptions of some methodological details, such as the generation of the randomisation

  14. Formin' actin in the nucleus.

    Science.gov (United States)

    Baarlink, Christian; Grosse, Robert

    2014-01-01

    Many if not most proteins can, under certain conditions, change cellular compartments, such as, for example, shuttling from the cytoplasm to the nucleus. Thus, many proteins may exert functions in various and very different subcellular locations, depending on the signaling context. A large amount of actin regulatory proteins has been detected in the mammalian cell nucleus, although their potential roles are much debated and are just beginning to emerge. Recently, members of the formin family of actin nucleators were also reported to dynamically localize to the nuclear environment. Here we discuss our findings that specific diaphanous-related formins can promote nuclear actin assembly in a signal-dependent manner.

  15. Superficie

    OpenAIRE

    Corna, Pablo María

    2015-01-01

    En la época posclásica del derecho romano se buscó, por parte del Estado y ciudadanos, utilizar ciertos terrenos que formaban parte de su patrimonio para arrendarlos por largo término —y a veces a perpetuidad— a personas que tenían el derecho de edificar o plantar mediante el pago de un precio llamado pensio o solárium. En el siglo II de nuestra era ya era utilizado para asentar tribus romanizadas en la frontera del imperio como para el cultivo de vid y los olivos que demoran varios años e...

  16. Chemotaxis and Actin Oscillations

    Science.gov (United States)

    Bodenschatz, Eberhard; Hsu, Hsin-Fang; Negrete, Jose; Beta, Carsten; Pumir, Alain; Gholami, Azam; Tarantola, Marco; Westendorf, Christian; Zykov, Vladimir

    Recently, self-oscillations of the cytoskeletal actin have been observed in Dictyostelium, a model system for studying chemotaxis. Here we report experimental results on the self-oscillation mechanism and the role of regulatory proteins and myosin II. We stimulate cells rapidly and periodically by using photo un-caging of the chemoattractant in a micro-fluidic device and measured the cellular responses. We found that the response amplitude grows with stimulation strength only in a very narrow region of stimulation, after which the response amplitude reaches a plateau. Moreover, the frequency-response is not constant but rather varies with the strength of external stimuli. To understand the underlying mechanism, we analyzed the polymerization and de-polymerization time in the single cell level. Despite of the large cell-to-cell variability, we found that the polymerization time is independent of external stimuli and the de-polymerization time is prolonged as the stimulation strength increases. Our conclusions will be summarized and the role of noise in the signaling network will be discussed. German Science Foundation CRC 937.

  17. Ring closure in actin polymers

    Energy Technology Data Exchange (ETDEWEB)

    Sinha, Supurna, E-mail: supurna@rri.res.in [Raman Research Institute, Bangalore 560080 (India); Chattopadhyay, Sebanti [Doon University, Dehradun 248001 (India)

    2017-03-18

    We present an analysis for the ring closure probability of semiflexible polymers within the pure bend Worm Like Chain (WLC) model. The ring closure probability predicted from our analysis can be tested against fluorescent actin cyclization experiments. We also discuss the effect of ring closure on bend angle fluctuations in actin polymers. - Highlights: • Ring closure of biopolymers. • Worm like chain model. • Predictions for experiments.

  18. Boolean gates on actin filaments

    International Nuclear Information System (INIS)

    Siccardi, Stefano; Tuszynski, Jack A.; Adamatzky, Andrew

    2016-01-01

    Actin is a globular protein which forms long polar filaments in the eukaryotic cytoskeleton. Actin networks play a key role in cell mechanics and cell motility. They have also been implicated in information transmission and processing, memory and learning in neuronal cells. The actin filaments have been shown to support propagation of voltage pulses. Here we apply a coupled nonlinear transmission line model of actin filaments to study interactions between voltage pulses. To represent digital information we assign a logical TRUTH value to the presence of a voltage pulse in a given location of the actin filament, and FALSE to the pulse's absence, so that information flows along the filament with pulse transmission. When two pulses, representing Boolean values of input variables, interact, then they can facilitate or inhibit further propagation of each other. We explore this phenomenon to construct Boolean logical gates and a one-bit half-adder with interacting voltage pulses. We discuss implications of these findings on cellular process and technological applications. - Highlights: • We simulate interaction between voltage pulses using on actin filaments. • We use a coupled nonlinear transmission line model. • We design Boolean logical gates via interactions between the voltage pulses. • We construct one-bit half-adder with interacting voltage pulses.

  19. Boolean gates on actin filaments

    Energy Technology Data Exchange (ETDEWEB)

    Siccardi, Stefano, E-mail: ssiccardi@2ssas.it [The Unconventional Computing Centre, University of the West of England, Bristol (United Kingdom); Tuszynski, Jack A., E-mail: jackt@ualberta.ca [Department of Oncology, University of Alberta, Edmonton, Alberta (Canada); Adamatzky, Andrew, E-mail: andrew.adamatzky@uwe.ac.uk [The Unconventional Computing Centre, University of the West of England, Bristol (United Kingdom)

    2016-01-08

    Actin is a globular protein which forms long polar filaments in the eukaryotic cytoskeleton. Actin networks play a key role in cell mechanics and cell motility. They have also been implicated in information transmission and processing, memory and learning in neuronal cells. The actin filaments have been shown to support propagation of voltage pulses. Here we apply a coupled nonlinear transmission line model of actin filaments to study interactions between voltage pulses. To represent digital information we assign a logical TRUTH value to the presence of a voltage pulse in a given location of the actin filament, and FALSE to the pulse's absence, so that information flows along the filament with pulse transmission. When two pulses, representing Boolean values of input variables, interact, then they can facilitate or inhibit further propagation of each other. We explore this phenomenon to construct Boolean logical gates and a one-bit half-adder with interacting voltage pulses. We discuss implications of these findings on cellular process and technological applications. - Highlights: • We simulate interaction between voltage pulses using on actin filaments. • We use a coupled nonlinear transmission line model. • We design Boolean logical gates via interactions between the voltage pulses. • We construct one-bit half-adder with interacting voltage pulses.

  20. Esophagectomy for Superficial Esophageal Neoplasia.

    Science.gov (United States)

    Watson, Thomas J

    2017-07-01

    Endoscopic therapies have become the standard of care for most cases of Barrett's esophagus with high-grade dysplasia or intramucosal adenocarcinoma. Despite a rapid and dramatic evolution in treatment paradigms, esophagectomy continues to occupy a place in the therapeutic armamentarium for superficial esophageal neoplasia. The managing physician must remain cognizant of the limitations of endoscopic approaches and consider surgical resection when they are exceeded. Esophagectomy, performed at experienced centers for appropriately selected patients with early-stage disease can be undertaken with the expectation of cure as well as low mortality, acceptable morbidity, and good long-term quality of life. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. Bacterial Actins? An Evolutionary Perspective

    Science.gov (United States)

    Doolittle, Russell F.; York, Amanda L.

    2003-01-01

    According to the conventional wisdom, the existence of a cytoskeleton in eukaryotes and its absence in prokaryotes constitute a fundamental divide between the two domains of life. An integral part of the dogma is that a cytoskeleton enabled an early eukaryote to feed upon prokaryotes, a consequence of which was the occasional endosymbiosis and the eventual evolution of organelles. Two recent papers present compelling evidence that actin, one of the principal components of a cytoskeleton, has a homolog in Bacteria that behaves in many ways like eukaryotic actin. Sequence comparisons reveml that eukaryotic actin and the bacterial homolog (mreB protein), unlike many other proteins common to eukaryotes and Bacteria, have very different and more highly extended evolutionary histories.

  2. Dendritic Actin Cytoskeleton: Structure, Functions, and Regulations

    Directory of Open Access Journals (Sweden)

    Anja Konietzny

    2017-05-01

    Full Text Available Actin is a versatile and ubiquitous cytoskeletal protein that plays a major role in both the establishment and the maintenance of neuronal polarity. For a long time, the most prominent roles that were attributed to actin in neurons were the movement of growth cones, polarized cargo sorting at the axon initial segment, and the dynamic plasticity of dendritic spines, since those compartments contain large accumulations of actin filaments (F-actin that can be readily visualized using electron- and fluorescence microscopy. With the development of super-resolution microscopy in the past few years, previously unknown structures of the actin cytoskeleton have been uncovered: a periodic lattice consisting of actin and spectrin seems to pervade not only the whole axon, but also dendrites and even the necks of dendritic spines. Apart from that striking feature, patches of F-actin and deep actin filament bundles have been described along the lengths of neurites. So far, research has been focused on the specific roles of actin in the axon, while it is becoming more and more apparent that in the dendrite, actin is not only confined to dendritic spines, but serves many additional and important functions. In this review, we focus on recent developments regarding the role of actin in dendrite morphology, the regulation of actin dynamics by internal and external factors, and the role of F-actin in dendritic protein trafficking.

  3. Geometrical Determinants of Neuronal Actin Waves

    OpenAIRE

    Tomba, Caterina; Bra?ni, C?line; Bugnicourt, Ghislain; Cohen, Floriane; Friedrich, Benjamin M.; Gov, Nir S.; Villard, Catherine

    2017-01-01

    Hippocampal neurons produce in their early stages of growth propagative, actin-rich dynamical structures called actin waves. The directional motion of actin waves from the soma to the tip of neuronal extensions has been associated with net forward growth, and ultimately with the specification of neurites into axon and dendrites. Here, geometrical cues are used to control actin wave dynamics by constraining neurons on adhesive stripes of various widths. A key observable, the average time betwe...

  4. Case for diagnosis. Actinic prurigo.

    Science.gov (United States)

    Daldon, Patricia Erica Christofoletti; Pascini, Mirella; Correa, Mariane

    2010-01-01

    A 13-year-old black boy had pruritic papular and nodular lesions on his forearms associated to edema of the lower lip, photophobia, conjunctivitis and pterygium. Skin biopsy of the lower lip revealed acanthosis, spongiosis with dermal perivascular mononuclear cell infiltration composed by lymphocytes, plasma cells and eosinophils consistent with actinic prurigo. Lesions improved considerably with the use of thalidomide 100mg/ day.

  5. Transforming growth factor β-induced superficial zone protein accumulation in the surface zone of articular cartilage is dependent on the cytoskeleton.

    Science.gov (United States)

    McNary, Sean M; Athanasiou, Kyriacos A; Reddi, A Hari

    2014-03-01

    The phenotype of articular chondrocytes is dependent on the cytoskeleton, specifically the actin microfilament architecture. Articular chondrocytes in monolayer culture undergo dedifferentiation and assume a fibroblastic phenotype. This process can be reversed by altering the actin cytoskeleton by treatment with cytochalasin. Whereas dedifferentiation has been studied on chondrocytes isolated from the whole cartilage, the effects of cytoskeletal alteration on specific zones of cells such as superficial zone chondrocytes are not known. Chondrocytes from the superficial zone secrete superficial zone protein (SZP), a lubricating proteoglycan that reduces the coefficient of friction of articular cartilage. A better understanding of this phenomenon may be useful in elucidating chondrocyte dedifferentiation in monolayer and accumulation of the cartilage lubricant SZP, with an eye toward tissue engineering functional articular cartilage. In this investigation, the effects of cytoskeletal modulation on the ability of superficial zone chondrocytes to secrete SZP were examined. Primary superficial zone chondrocytes were cultured in monolayer and treated with a combination of cytoskeleton modifying reagents and transforming growth factor β (TGFβ) 1, a critical regulator of SZP production. Whereas cytochalasin D maintains the articular chondrocyte phenotype, the hallmark of the superficial zone chondrocyte, SZP, was inhibited in the presence of TGFβ1. A decrease in TGFβ1-induced SZP accumulation was also observed when the microtubule cytoskeleton was modified using paclitaxel. These effects of actin and microtubule alteration were confirmed through the application of jasplakinolide and colchicine, respectively. As Rho GTPases regulate actin organization and microtubule polymerization, we hypothesized that the cytoskeleton is critical for TGFβ-induced SZP accumulation. TGFβ-mediated SZP accumulation was inhibited by small molecule inhibitors ML141 (Cdc42), NSC23766 (Rac1

  6. Effects of F/G-actin ratio and actin turn-over rate on NADPH oxidase activity in microglia

    DEFF Research Database (Denmark)

    Rasmussen, Izabela; Pedersen, Line Hjortshøj; Byg, Luise

    2010-01-01

    Most in vivo studies that have addressed the role of actin dynamics in NADPH oxidase function in phagocytes have used toxins to modulate the polymerization state of actin and mostly effects on actin has been evaluated by end point measurements of filamentous actin, which says little about actin d...... dynamics, and without consideration for the subcellular distribution of the perturbed actin cytoskeleton....

  7. The actin multigene family of Paramecium tetraurelia

    Directory of Open Access Journals (Sweden)

    Wagner Erika

    2007-03-01

    Full Text Available Abstract Background A Paramecium tetraurelia pilot genome project, the subsequent sequencing of a Megabase chromosome as well as the Paramecium genome project aimed at gaining insight into the genome of Paramecium. These cells display a most elaborate membrane trafficking system, with distinct, predictable pathways in which actin could participate. Previously we had localized actin in Paramecium; however, none of the efforts so far could proof the occurrence of actin in the cleavage furrow of a dividing cell, despite the fact that actin is unequivocally involved in cell division. This gave a first hint that Paramecium may possess actin isoforms with unusual characteristics. The genome project gave us the chance to search the whole Paramecium genome, and, thus, to identify and characterize probably all actin isoforms in Paramecium. Results The ciliated protozoan, P. tetraurelia, contains an actin multigene family with at least 30 members encoding actin, actin-related and actin-like proteins. They group into twelve subfamilies; a large subfamily with 10 genes, seven pairs and one trio with > 82% amino acid identity, as well as three single genes. The different subfamilies are very distinct from each other. In comparison to actins in other organisms, P. tetraurelia actins are highly divergent, with identities topping 80% and falling to 30%. We analyzed their structure on nucleotide level regarding the number and position of introns. On amino acid level, we scanned the sequences for the presence of actin consensus regions, for amino acids of the intermonomer interface in filaments, for residues contributing to ATP binding, and for known binding sites for myosin and actin-specific drugs. Several of those characteristics are lacking in several subfamilies. The divergence of P. tetraurelia actins and actin-related proteins between different P. tetraurelia subfamilies as well as with sequences of other organisms is well represented in a phylogenetic

  8. Mesoscopic model of actin-based propulsion.

    Directory of Open Access Journals (Sweden)

    Jie Zhu

    Full Text Available Two theoretical models dominate current understanding of actin-based propulsion: microscopic polymerization ratchet model predicts that growing and writhing actin filaments generate forces and movements, while macroscopic elastic propulsion model suggests that deformation and stress of growing actin gel are responsible for the propulsion. We examine both experimentally and computationally the 2D movement of ellipsoidal beads propelled by actin tails and show that neither of the two models can explain the observed bistability of the orientation of the beads. To explain the data, we develop a 2D hybrid mesoscopic model by reconciling these two models such that individual actin filaments undergoing nucleation, elongation, attachment, detachment and capping are embedded into the boundary of a node-spring viscoelastic network representing the macroscopic actin gel. Stochastic simulations of this 'in silico' actin network show that the combined effects of the macroscopic elastic deformation and microscopic ratchets can explain the observed bistable orientation of the actin-propelled ellipsoidal beads. To test the theory further, we analyze observed distribution of the curvatures of the trajectories and show that the hybrid model's predictions fit the data. Finally, we demonstrate that the model can explain both concave-up and concave-down force-velocity relations for growing actin networks depending on the characteristic time scale and network recoil. To summarize, we propose that both microscopic polymerization ratchets and macroscopic stresses of the deformable actin network are responsible for the force and movement generation.

  9. LL-37 induces polymerization and bundling of actin and affects actin structure.

    Directory of Open Access Journals (Sweden)

    Asaf Sol

    Full Text Available Actin exists as a monomer (G-actin which can be polymerized to filaments F-actin that under the influence of actin-binding proteins and polycations bundle and contribute to the formation of the cytoskeleton. Bundled actin from lysed cells increases the viscosity of sputum in lungs of cystic fibrosis patients. The human host defense peptide LL-37 was previously shown to induce actin bundling and was thus hypothesized to contribute to the pathogenicity of this disease. In this work, interactions between actin and the cationic LL-37 were studied by optical, proteolytic and surface plasmon resonance methods and compared to those obtained with scrambled LL-37 and with the cationic protein lysozyme. We show that LL-37 binds strongly to CaATP-G-actin while scrambled LL-37 does not. While LL-37, at superstoichiometric LL-37/actin concentrations polymerizes MgATP-G-actin, at lower non-polymerizing concentrations LL-37 inhibits actin polymerization by MgCl(2 or NaCl. LL-37 bundles Mg-F-actin filaments both at low and physiological ionic strength when in equimolar or higher concentrations than those of actin. The LL-37 induced bundles are significantly less sensitive to increase in ionic strength than those induced by scrambled LL-37 and lysozyme. LL-37 in concentrations lower than those needed for actin polymerization or bundling, accelerates cleavage of both monomer and polymer actin by subtilisin. Our results indicate that the LL-37-actin interaction is partially electrostatic and partially hydrophobic and that a specific actin binding sequence in the peptide is responsible for the hydrophobic interaction. LL-37-induced bundles, which may contribute to the accumulation of sputum in cystic fibrosis, are dissociated very efficiently by DNase-1 and also by cofilin.

  10. Scanning electron microscopy of superficial white onychomycosis*

    Science.gov (United States)

    de Almeida Jr., Hiram Larangeira; Boabaid, Roberta Oliveira; Timm, Vitor; Silva, Ricardo Marques e; de Castro, Luis Antonio Suita

    2015-01-01

    Superficial white onychomycosis is characterized by opaque, friable, whitish superficial spots on the nail plate. We examined an affected halux nail of a 20-year-old male patient with scanning electron microscopy. The mycological examination isolated Trichophyton mentagrophytes. Abundant hyphae with the formation of arthrospores were found on the nail's surface, forming small fungal colonies. These findings showed the great capacity for dissemination of this form of onychomycosis. PMID:26560225

  11. Probing friction in actin-based motility

    International Nuclear Information System (INIS)

    Marcy, Yann; Joanny, Jean-Francois; Prost, Jacques; Sykes, Cecile

    2007-01-01

    Actin dynamics are responsible for cell protrusion and certain intracellular movements. The transient attachment of the actin filaments to a moving surface generates a friction force that resists the movement. We probe here the dynamics of these attachments by inducing a stick-slip behavior via micromanipulation of a growing actin comet. We show that general principles of adhesion and friction can explain our observations

  12. Cytoskeletal actin dynamics shape a ramifying actin network underpinning immunological synapse formation

    DEFF Research Database (Denmark)

    Fritzsche, Marco; Fernandes, Ricardo A.; Chang, Veronica T.

    2017-01-01

    optical microscopes to analyze resting and activated T cells, we show that, following contact formation with activating surfaces, these cells sequentially rearrange their cortical actin across the entire cell, creating a previously unreported ramifying actin network above the immunological synapse...

  13. Superficial chemical peeling with salicylic acid in facial dermatoses

    International Nuclear Information System (INIS)

    Bari, A.U.; Iqbal, Z.; Rahman, S.B.

    2007-01-01

    To determine the effectiveness of salicylic acid chemical peeling in common dermatological conditions affecting face in people with predominant Fitzpatrick skin type IV and V. A total of 167 patients of either gender, aged between 13 to 60 years, having some facial dermatoses (melasma, acne vulgaris, postinflammatory hyperpigmentations, freckles, fine lines and wrinkles, post-inflammatory scars, actinic keratoses, and plane facial warts) were included. A series of eight weekly hospital based peeling sessions was conducted in all patients under standardized conditions with 30% salicylic acid. Clinical improvement in different disorders was evaluated by change in MASI score, decrease in the size of affected area and % reduction in lesions count. McNemar test was applied for data analysis. Majority of the patients showed moderate to excellent response. There was 35% to 63% improvement (p< 0.05) in all dermatoses. Significant side effects, as feared in Asian skins were not observed. Chemical peeling with salicylic acid is an effective and safe treatment modality in many superficial facial dermatoses. (author)

  14. Bioinformatics study of the mangrove actin genes

    Science.gov (United States)

    Basyuni, M.; Wasilah, M.; Sumardi

    2017-01-01

    This study describes the bioinformatics methods to analyze eight actin genes from mangrove plants on DDBJ/EMBL/GenBank as well as predicted the structure, composition, subcellular localization, similarity, and phylogenetic. The physical and chemical properties of eight mangroves showed variation among the genes. The percentage of the secondary structure of eight mangrove actin genes followed the order of a helix > random coil > extended chain structure for BgActl, KcActl, RsActl, and A. corniculatum Act. In contrast to this observation, the remaining actin genes were random coil > extended chain structure > a helix. This study, therefore, shown the prediction of secondary structure was performed for necessary structural information. The values of chloroplast or signal peptide or mitochondrial target were too small, indicated that no chloroplast or mitochondrial transit peptide or signal peptide of secretion pathway in mangrove actin genes. These results suggested the importance of understanding the diversity and functional of properties of the different amino acids in mangrove actin genes. To clarify the relationship among the mangrove actin gene, a phylogenetic tree was constructed. Three groups of mangrove actin genes were formed, the first group contains B. gymnorrhiza BgAct and R. stylosa RsActl. The second cluster which consists of 5 actin genes the largest group, and the last branch consist of one gene, B. sexagula Act. The present study, therefore, supported the previous results that plant actin genes form distinct clusters in the tree.

  15. Geometrical Determinants of Neuronal Actin Waves.

    Science.gov (United States)

    Tomba, Caterina; Braïni, Céline; Bugnicourt, Ghislain; Cohen, Floriane; Friedrich, Benjamin M; Gov, Nir S; Villard, Catherine

    2017-01-01

    Hippocampal neurons produce in their early stages of growth propagative, actin-rich dynamical structures called actin waves. The directional motion of actin waves from the soma to the tip of neuronal extensions has been associated with net forward growth, and ultimately with the specification of neurites into axon and dendrites. Here, geometrical cues are used to control actin wave dynamics by constraining neurons on adhesive stripes of various widths. A key observable, the average time between the production of consecutive actin waves, or mean inter-wave interval (IWI), was identified. It scales with the neurite width, and more precisely with the width of the proximal segment close to the soma. In addition, the IWI is independent of the total number of neurites. These two results suggest a mechanistic model of actin wave production, by which the material conveyed by actin waves is assembled in the soma until it reaches the threshold leading to the initiation and propagation of a new actin wave. Based on these observations, we formulate a predictive theoretical description of actin wave-driven neuronal growth and polarization, which consistently accounts for different sets of experiments.

  16. Pharmacological treatment of actinic keratosis

    Directory of Open Access Journals (Sweden)

    Ewa Zwierzyńska

    2016-09-01

    Full Text Available Actinic keratosis (AK is a disease characterized by hyperkeratotic lesions on skin damaged by ultraviolet. radiation. These lesions may progress to squamous cell or basal cell carcinoma. Currently pharmacotherapy and different surgical procedures are used in AK therapy. The most common treatment options are 5-fluorouracil, imiquimod, diclofenac, ingenol mebutate, and first and third generation retinoids (retinol, adapalene, tazarotene. Furthermore, research is being carried out in order to test new medications including nicotinamide, resiquimod, piroxicam, potassium dobesilate and oleogel based on a triterpene extract (betulin, betulinic acid. Recently, the preventive effect of acetylsalicylic acid and celecoxib has also been investigated.

  17. Unconventional actin conformations localize on intermediate filaments in mitosis

    International Nuclear Information System (INIS)

    Hubert, Thomas; Vandekerckhove, Joel; Gettemans, Jan

    2011-01-01

    Research highlights: → Unconventional actin conformations colocalize with vimentin on a cage-like structure in metaphase HEK 293T cells. → These conformations are detected with the anti-actin antibodies 1C7 ('lower dimer') and 2G2 ('nuclear actin'), but not C4 (monomeric actin). → Mitotic unconventional actin cables are independent of filamentous actin or microtubules. → Unconventional actin colocalizes with vimentin on a nocodazole-induced perinuclear dense mass of cables. -- Abstract: Different structural conformations of actin have been identified in cells and shown to reside in distinct subcellular locations of cells. In this report, we describe the localization of actin on a cage-like structure in metaphase HEK 293T cells. Actin was detected with the anti-actin antibodies 1C7 and 2G2, but not with the anti-actin antibody C4. Actin contained in this structure is independent of microtubules and actin filaments, and colocalizes with vimentin. Taking advantage of intermediate filament collapse into a perinuclear dense mass of cables when microtubules are depolymerized, we were able to relocalize actin to such structures. We hypothesize that phosphorylation of intermediate filaments at mitosis entry triggers the recruitment of different actin conformations to mitotic intermediate filaments. Storage and partition of the nuclear actin and antiparallel 'lower dimer' actin conformations between daughter cells possibly contribute to gene transcription and transient actin filament dynamics at G1 entry.

  18. Dynamic Regulation of Sarcomeric Actin Filaments in Striated Muscle

    OpenAIRE

    Ono, Shoichiro

    2010-01-01

    In striated muscle, the actin cytoskeleton is differentiated into myofibrils. Actin and myosin filaments are organized in sarcomeres and specialized for producing contractile forces. Regular arrangement of actin filaments with uniform length and polarity is critical for the contractile function. However, the mechanisms of assembly and maintenance of sarcomeric actin filaments in striated muscle are not completely understood. Live imaging of actin in striated muscle has revealed that actin sub...

  19. Actin expression in some Platyhelminthe species.

    Science.gov (United States)

    Fagotti, A; Panara, F; Di Rosa, I; Simoncelli, F; Gabbiani, G; Pascolini, R

    1994-10-01

    Actin expression in some Platyhelminthe species was demonstrated by western-blotting and immunocytochemical analysis using two distinct anti-actin antibodies: the anti-total actin that reacts against all actin isoforms of higher vertebrates and the anti-alpha SM-1 that recognizes the alpha-smooth muscle (alpha SM) isotype of endothermic vertebrates (Skalli et al., 1986). Western-blotting experiments showed that all species tested, including some free-living Platyhelminthes (Tricladida and Rhabdocoela) and the parasitic Fasciola hepatica, were stained by anti-total actin antibody while only Dugesidae and Dendrocoelidae showed a positive immunoreactivity against anti-alpha SM-1. These results were confirmed by cytochemical immunolocalization using both avidin biotin conjugated peroxidase reaction on paraffin sections, and immunogold staining on Lowicryl 4KM embedded specimens. Our findings may contribute to the understanding of Platyhelminthes phylogeny.

  20. Tolerance and safety of superficial chemical peeling with salicylic acid in various facial dermatoses

    Directory of Open Access Journals (Sweden)

    Iqbal Zafar

    2005-03-01

    Full Text Available BACKGROUND: Chemical peeling is a skin-wounding procedure that may have some potentially undesirable side-effects. AIMS: The present study is directed towards safety concerns associated with superficial chemical peeling with salicylic acid in various facial dermatoses. METHODS: The study was a non-comparative and a prospective one. Two hundred and sixty-eight patients of either sex, aged between 10 to 60 years, undergoing superficial chemical peeling for various facial dermatoses (melasma, acne vulgaris, freckles, post-inflammatory scars/pigmentation, actinic keratoses, plane facial warts, etc. were included in the study. Eight weekly peeling sessions were carried out in each patient. Tolerance to the procedure and any undesirable effects noted during these sessions were recorded. RESULTS: Almost all the patients tolerated the procedure well. Mild discomfort, burning, irritation and erythema were quite common but the incidence of major side-effects was very low and these too, were easily manageable. There was no significant difference in the incidence of side-effects between facial dermatoses (melasma, acne and other pigmentary disorders. CONCLUSION: Chemical peeling with salicylic acid is a well tolerated and safe treatment modality in many superficial facial dermatoses.

  1. Managing actinic keratosis in primary care.

    Science.gov (United States)

    Salmon, Nicola; Tidman, Michael J

    2016-10-01

    Actinic, or solar, keratosis is caused by chronic ultraviolet-induced damage to the epidermis. In the UK, 15-23% of individuals have actinic keratosis lesions. Risk factors include: advanced age; male gender; cumulative sun exposure or phototherapy; Fitzpatrick skin phototypes I-II; long-term immuno-suppression and genetic syndromes e.g. xeroderma pigmentosum and albinism. Actinic keratoses are regarded by some authorities as premalignant lesions that may transform into invasive squamous cell carcinoma (SCC) and by others as in situ SCC that may progress to an invasive stage. The risk of malignant change appears low; up to 0.5% per lesion per year. Up to 20-30% of lesions may spontaneously regress but in the absence of any reliable prognostic clinical indicators regarding malignant potential active treatment is considered appropriate. Actinic keratosis lesions may present as discrete hyperkeratotic papules, cutaneous horns, or more subtle flat lesions on sun-exposed areas of skin. The single most helpful diagnostic sign is an irregularly roughened surface texture: a sandpaper-like feel almost always indicates actinic damage. Dermatoscopy can be helpful in excluding signs of basal cell carcinoma when actinic keratosis is non-keratotic. It is always important to consider the possibility of SCC. The principal indication for referral to secondary care is the possibility of cutaneous malignancy. However, widespread and severe actinic damage in patients who are immunosuppressed is also a reason for referral.

  2. HIGH ORIGIN OF SUPERFICIAL ULNAR ARTERY- A CASE REPORT

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    Anjana Jayakumaran Nair

    2017-03-01

    Full Text Available BACKGROUND High origin and superficially placed ulnar artery is a rare anatomical variant that usually arises either in the axilla or arm and runs a superficial course in the forearm, enters the hand and participates in the formation of superficial palmar arch. During routine dissection of cadavers in our department, we observed a unilateral case of high origin and superficial ulnar artery in a human male cadaver. It originated from the brachial artery in the lower third of arm 4 cm above its bifurcation. From its origin, it passed downwards along the medial aspect of forearm, superficial to the flexors, entered hand superficial to the flexor retinaculum and formed superficial palmar arch. The knowledge of existence of a superficial ulnar artery is important during vascular and reconstructive surgery and also in evaluation of angiographic images. Superficial position makes it more vulnerable to trauma and more accessible to cannulation.

  3. Intravesical Gemcitabine for Treatment of Superficial Bladder ...

    African Journals Online (AJOL)

    Objectives: Intravesical Bacillus Calmette-Guérin (BCG) vaccine is the mainstay of treatment and prophylaxis in superficial bladder cancer (SBC) as it reduces tumor recurrence and disease progression. About one-third of patients do not respond to BCG. The aim of this study was to determine the efficacy of intravesical ...

  4. Actinic Granuloma with Focal Segmental Glomerulosclerosis

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    Ruedee Phasukthaworn

    2016-02-01

    Full Text Available Actinic granuloma is an uncommon granulomatous disease, characterized by annular erythematous plaque with central clearing predominately located on sun-damaged skin. The pathogenesis is not well understood, ultraviolet radiation is recognized as precipitating factor. We report a case of a 52-year-old woman who presented with asymptomatic annular erythematous plaques on the forehead and both cheeks persisting for 2 years. The clinical presentation and histopathologic findings support the diagnosis of actinic granuloma. During that period of time, she also developed focal segmental glomerulosclerosis. The association between actinic granuloma and focal segmental glomerulosclerosis needs to be clarified by further studies.

  5. [Superficial venous thrombosis. A state of art].

    Science.gov (United States)

    Sándor, Tamás

    2017-01-01

    For a long time superficial thrombophlebitis has been thought to be a rather benign condition. Recently, when duplex ultrasound technique is used for the diagnosis more and more often, the disease is proved to be more dangerous than anticipated. Thrombosis propagates to the deep veins in 6-44% and pulmonary embolism was observed on the patients in 1,5-33%. We can calculate venous thromboembolic complications on every fourth patient. Diagnosis is clinical, but duplex ultrasound examination is mandatory, for estimation of the thrombus extent, for exclusion of the deep venous thrombosis and for follow up. Both legs should be checked with ultrasound, because simultaneous deep venous thrombosis can develop on the contralateral limb. Two different forms can be distinguished: superficial venous thrombosis with, or without varicose veins. In cases of spontaneous, non varicous form, especially when the process is migrating or recurrent, a careful clinical examination is necessery for exclusion of malignant diseases and thrombophilia. The treatment options are summarised on the basis of recent international consensus statements. The American and German guidelines are similar. Compression and mobilisation are cornerstones of the therapy. For a short segment thrombosis non steroidal antiinflammatory drugs are effective. For longer segments low molecular-weight heparins are preferred. Information on the effect of the novel oral anticoagulants for the therapy is lacking but they may appear to be effective in the future for this indication. When thrombus is close to the sapheno-femoral or sapheno-popliteal junction crossectomy (high ligation), or low molecular-weight heparin in therapeutic doses are indicated. The term superficial thrombophlebitis should be discouraged, because inflammation and infection is not the primary pathology. It should be called correctly superficial venous thrombosis in order to avoid the unnecessary administration of antibiotics and the misconception

  6. Mechanics model for actin-based motility.

    Science.gov (United States)

    Lin, Yuan

    2009-02-01

    We present here a mechanics model for the force generation by actin polymerization. The possible adhesions between the actin filaments and the load surface, as well as the nucleation and capping of filament tips, are included in this model on top of the well-known elastic Brownian ratchet formulation. A closed form solution is provided from which the force-velocity relationship, summarizing the mechanics of polymerization, can be drawn. Model predictions on the velocity of moving beads driven by actin polymerization are consistent with experiment observations. This model also seems capable of explaining the enhanced actin-based motility of Listeria monocytogenes and beads by the presence of Vasodilator-stimulated phosphoprotein, as observed in recent experiments.

  7. Malignant transformation of superficial peritoneal endometriosis lesion.

    Science.gov (United States)

    Marchand, Eva; Hequet, Delphine; Thoury, Anne; Barranger, Emmanuel

    2013-08-26

    A 63-year-old woman with no medical history underwent an abdominal surgery with hysterectomy and bilateral salpingo-oophorectomy for a 10 cm peritoneal cyst with increased cancer antigene-125. A large suspicious tumour of the Douglas space, with contact to the uterus and the rectal wall was described. The rest of the exploration was normal, specially the rest of the peritoneum. Histopathology revealed a malignant transformation of a superficial peritoneal endometriosis. Secondary surgery was thus completed by laparoscopy with bilateral pelvic and para-aortic lymph node dissections, omentectomy and multiple peritoneal biopsies. All staging samples were free of cancer; therefore no complementary therapy was administered. After 18 months of follow-up, consisting of clinical examination and pelvis magnetic resonance imaging every 6 months, we did not observe any recurrence. Malignant transformation of superficial peritoneal endometriosis is a rare disease and surgical management seems to be the main treatment.

  8. Clinico Mycological Study of Superficial Mycoses

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    Rachana J. Magdum

    2016-10-01

    Full Text Available Background: Generally it is well established fact that geographical distribution of the fungi may change from time to time; hence this study was planned. Aim and Objectives: To analyze the prevalence of superficial mycoses, its clinical presentation and species identification of the fungal isolates responsible for the disease. Material and Methods: A total 125 clinically diagnosed cases of superficial mycoses visiting Dermatology and Venereology outpatient department of Bharati Hospital, Sangli, for a period of one year were included in the study. Specimens like skin scrapping, nail clipping, hair were collected and subjected to KOH mount and culture. Identification of species was done by macroscopic examination of culture, tease mount and other physiological tests including Urease test, Hair perforation tests and Germ tube test. Results: Superficial mycosis was more common in the age group of 21-30 years (28% and in males (60.8%. The infection was more common in students (29.6%. Tinea corporis (42.4% was the commonest clinical type followed by tinea cruris (22.4%. 61.6% cases were positive by direct microscopy and 60.8% cases showed culture positive. Out of 125 samples, dermatophytes were grown in 63 cases (82.89% followed by non dermatophytic moulds in 10 cases (13.16% and Candida albicans in 3 cases (3.95%. The most common isolate among dermtophytosis was T. rubrum (46.05% followed by T. mentagrophyte (25%. Conclusion: It was concluded that along with dermatophytes, non dermatophytic moulds are also important to cause of superficial mycoses

  9. Spectral characterization of superficial coal groups

    International Nuclear Information System (INIS)

    Ahmad, I.; Khan, M.A.; Ishaq, M.; Shakirullah; Bahadur, A.

    2004-01-01

    Spectral characterization of superficial coal groups was performed in KBr pellets. KBr Pellets were prepared for virgin and variously pretreated coal samples. Spectra of satisfactory resolution were obtained in wave number range-4000-400 cm /sup -1/. Presence of broad absorption bands corresponds to hydroxyl, carbonyl, carboxyl and phenolic functionalities in the spectra clearly define their presence in all samples understudy. Forced oxidation proved effective for oxidation of both aliphatic and aromatic configurations, which can be revealed from the respective spectra. (author)

  10. Volumetric and superficial characterization of carbon activated

    International Nuclear Information System (INIS)

    Carrera G, L.M.; Garcia S, I.; Jimenez B, J.; Solache R, M.; Lopez M, B.; Bulbulian G, S.; Olguin G, M.T.

    2000-01-01

    The activated carbon is the resultant material of the calcination process of natural carbonated materials as coconut shells or olive little bones. It is an excellent adsorbent of diluted substances, so much in colloidal form, as in particles form. Those substances are attracted and retained by the carbon surface. In this work is make the volumetric and superficial characterization of activated carbon treated thermically (300 Centigrade) in function of the grain size average. (Author)

  11. HISTOPATHOLOGY OF MARGINAL SUPERFICIAL PERIODONTIUM AT MENOPAUSE

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    A. Georgescu

    2012-03-01

    Full Text Available Premises: Sexual hormones may affect the general health condition of women, as early as puberty, continuing during pregnancy and also after menopause. Variations of the hormonal levels may cause different – either local or general – pathological modifications. Sexual hormones may also affect periodontal status, favourizing gingival inflammations and reducing periodontal resistance to the action of the bacterial plaque. Scope: Establishment of the correlations between the debut or the manifestation of menopause and the modifications produced in the superficial periodontium. Materials and method: Clinical and paraclinical investigations were performed on female patients with ages between 45 and 66 years, involving macroscopic, microscopic and radiological recording of the aspect of the superificial periodontium (gingiva. Results: Analysis of the histological sections evidenced atrophic and involutive modifications in the marginal superficial periodontium of female patients at menopause. Conclusions: Sexual hormones intervene in the histological equilibrium of the marginal superficial periodontium, influencing the periodontal health status, which explains the correlation between the subjective symptomatology specific to menopause and the histopatological aspect at epithelial level.

  12. Diagnosis and treatment of superficial esophageal cancer.

    Science.gov (United States)

    Barret, Maximilien; Prat, Frédéric

    2018-01-01

    Endoscopy allows for the screening, early diagnosis, treatment and follow up of superficial esophageal cancer. Endoscopic submucosal dissection has become the gold standard for the resection of superficial squamous cell neoplasia. Combinations of endoscopic mucosal resection and radiofrequency ablation are the mainstay of the management of Barrett's associated neoplasia. However, protruded, non-lifting or large lesions may be better managed by endoscopic submucosal dissection. Novel ablation tools, such as argon plasma coagulation with submucosal lifting and cryoablation balloons, are being developed for the treatment of residual Barrett's esophagus, since iatrogenic strictures still hamper the development of extensive circumferential resections in the esophagus. Optimal surveillance modalities after endoscopic resection are still to be determined. The assessment of the risk of lymph-node metastases, as well as of the need for additional treatments based on qualitative and quantitative histological criteria, balanced to the patient's condition, requires a dedicated multidisciplinary team decision process. The need for trained endoscopists, expert pathologists and surgeons, and specialized multidisciplinary meetings underlines the role of expert centers in the management of superficial esophageal cancer.

  13. What factors control superficial lava dome explosivity?

    Science.gov (United States)

    Boudon, Georges; Balcone-Boissard, Hélène; Villemant, Benoît; Morgan, Daniel J

    2015-09-30

    Dome-forming eruption is a frequent eruptive style and a major hazard on numerous volcanoes worldwide. Lava domes are built by slow extrusion of degassed, viscous magma and may be destroyed by gravitational collapse or explosion. The triggering of lava dome explosions is poorly understood: here we propose a new model of superficial lava-dome explosivity based upon a textural and geochemical study (vesicularity, microcrystallinity, cristobalite distribution, residual water contents, crystal transit times) of clasts produced by key eruptions. Superficial explosion of a growing lava dome may be promoted through porosity reduction caused by both vesicle flattening due to gas escape and syn-eruptive cristobalite precipitation. Both processes generate an impermeable and rigid carapace allowing overpressurisation of the inner parts of the lava dome by the rapid input of vesiculated magma batches. The relative thickness of the cristobalite-rich carapace is an inverse function of the external lava dome surface area. Explosive activity is thus more likely to occur at the onset of lava dome extrusion, in agreement with observations, as the likelihood of superficial lava dome explosions depends inversely on lava dome volume. This new result is of interest for the whole volcanological community and for risk management.

  14. Separation of actin-dependent and actin-independent lipid rafts

    NARCIS (Netherlands)

    Klappe, Karin; Hummel, Ina; Kok, Jan Willem

    2013-01-01

    Lipid rafts have been isolated on the basis of their resistance to various detergents and more recently by using detergent-free procedures. The actin cytoskeleton is now recognized as a dynamic regulator of lipid raft stability. We carefully analyzed the effects of the cortical actin-disrupting

  15. Green fluorescent protein-mtalin causes defects in actin organization and cell expansion in Arabidopsis and inhibits actin depolymerizing factor's actin depolymerizing activity in vitro

    NARCIS (Netherlands)

    Ketelaar, T.; Anthony, R.G.; Hussey, P.J.

    2004-01-01

    Expression of green fluorescent protein (GFP) linked to an actin binding domain is a commonly used method for live cell imaging of the actin cytoskeleton. One of these chimeric proteins is GFP-mTalin (GFP fused to the actin binding domain of mouse talin). Although it has been demonstrated that

  16. Structural Basis of Actin Filament Nucleation by Tandem W Domains

    Science.gov (United States)

    Chen, Xiaorui; Ni, Fengyun; Tian, Xia; Kondrashkina, Elena; Wang, Qinghua; Ma, Jianpeng

    2013-01-01

    SUMMARY Spontaneous nucleation of actin is very inefficient in cells. To overcome this barrier, cells have evolved a set of actin filament nucleators to promote rapid nucleation and polymerization in response to specific stimuli. However, the molecular mechanism of actin nucleation remains poorly understood. This is hindered largely by the fact that actin nucleus, once formed, rapidly polymerizes into filament, thus making it impossible to capture stable multisubunit actin nucleus. Here, we report an effective double-mutant strategy to stabilize actin nucleus by preventing further polymerization. Employing this strategy, we solved the crystal structure of AMPPNP-actin in complex with the first two tandem W domains of Cordon-bleu (Cobl), a potent actin filament nucleator. Further sequence comparison and functional studies suggest that the nucleation mechanism of Cobl is probably shared by the p53 cofactor JMY, but not Spire. Moreover, the double-mutant strategy opens the way for atomic mechanistic study of actin nucleation and polymerization. PMID:23727244

  17. Incorporation of mammalian actin into microfilaments in plant cell nucleus

    Directory of Open Access Journals (Sweden)

    Paves Heiti

    2004-04-01

    Full Text Available Abstract Background Actin is an ancient molecule that shows more than 90% amino acid homology between mammalian and plant actins. The regions of the actin molecule that are involved in F-actin assembly are largely conserved, and it is likely that mammalian actin is able to incorporate into microfilaments in plant cells but there is no experimental evidence until now. Results Visualization of microfilaments in onion bulb scale epidermis cells by different techniques revealed that rhodamine-phalloidin stained F-actin besides cytoplasm also in the nuclei whereas GFP-mouse talin hybrid protein did not enter the nuclei. Microinjection of fluorescently labeled actin was applied to study the presence of nuclear microfilaments in plant cells. Ratio imaging of injected fluorescent rabbit skeletal muscle actin and phalloidin staining of the microinjected cells showed that mammalian actin was able to incorporate into plant F-actin. The incorporation occurred preferentially in the nucleus and in the perinuclear region of plant cells whereas part of plant microfilaments, mostly in the periphery of cytoplasm, did not incorporate mammalian actin. Conclusions Microinjected mammalian actin is able to enter plant cell's nucleus, whereas incorporation of mammalian actin into plant F-actin occurs preferentially in the nucleus and perinuclear area.

  18. The actinome of Dictyostelium discoideum in comparison to actins and actin-related proteins from other organisms.

    Directory of Open Access Journals (Sweden)

    Jayabalan M Joseph

    Full Text Available Actin belongs to the most abundant proteins in eukaryotic cells which harbor usually many conventional actin isoforms as well as actin-related proteins (Arps. To get an overview over the sometimes confusing multitude of actins and Arps, we analyzed the Dictyostelium discoideum actinome in detail and compared it with the genomes from other model organisms. The D. discoideum actinome comprises 41 actins and actin-related proteins. The genome contains 17 actin genes which most likely arose from consecutive gene duplications, are all active, in some cases developmentally regulated and coding for identical proteins (Act8-group. According to published data, the actin fraction in a D. discoideum cell consists of more than 95% of these Act8-type proteins. The other 16 actin isoforms contain a conventional actin motif profile as well but differ in their protein sequences. Seven actin genes are potential pseudogenes. A homology search of the human genome using the most typical D. discoideum actin (Act8 as query sequence finds the major actin isoforms such as cytoplasmic beta-actin as best hit. This suggests that the Act8-group represents a nearly perfect actin throughout evolution. Interestingly, limited data from D. fasciculatum, a more ancient member among the social amoebae, show different relationships between conventional actins. The Act8-type isoform is most conserved throughout evolution. Modeling of the putative structures suggests that the majority of the actin-related proteins is functionally unrelated to canonical actin. The data suggest that the other actin variants are not necessary for the cytoskeleton itself but rather regulators of its dynamical features or subunits in larger protein complexes.

  19. Xenopus egg cytoplasm with intact actin.

    Science.gov (United States)

    Field, Christine M; Nguyen, Phuong A; Ishihara, Keisuke; Groen, Aaron C; Mitchison, Timothy J

    2014-01-01

    We report optimized methods for preparing Xenopus egg extracts without cytochalasin D, that we term "actin-intact egg extract." These are undiluted egg cytoplasm that contains abundant organelles, and glycogen which supplies energy, and represents the least perturbed cell-free cytoplasm preparation we know of. We used this system to probe cell cycle regulation of actin and myosin-II dynamics (Field et al., 2011), and to reconstitute the large, interphase asters that organize early Xenopus embryos (Mitchison et al., 2012; Wühr, Tan, Parker, Detrich, & Mitchison, 2010). Actin-intact Xenopus egg extracts are useful for analysis of actin dynamics, and interaction of actin with other cytoplasmic systems, in a cell-free system that closely mimics egg physiology, and more generally for probing the biochemistry and biophysics of the egg, zygote, and early embryo. Detailed protocols are provided along with assays used to check cell cycle state and tips for handling and storing undiluted egg extracts. © 2014 Elsevier Inc. All rights reserved.

  20. Antibodies to actin in autoimmune haemolytic anaemia

    Directory of Open Access Journals (Sweden)

    Ritzmann Mathias

    2010-03-01

    Full Text Available Abstract Background In autoimmune haemolytic anaemia (AIHA, autoreactive antibodies directed against red blood cells are up-regulated, leading to erythrocyte death. Mycoplasma suis infections in pigs induce AIHA of both the warm and cold types. The aim of this study was to identify the target autoantigens of warm autoreactive IgG antibodies. Sera from experimentally M. suis-infected pigs were screened for autoreactivity. Results Actin-reactive antibodies were found in the sera of 95% of all animals tested. The reactivity was species-specific, i.e. reactivity with porcine actin was significantly higher than with rabbit actin. Sera of animals previously immunised with the M. suis adhesion protein MSG1 showed reactivity with actin prior to infection with M. suis indicating that molecular mimicry is involved in the specific autoreactive mechanism. A potentially cross-reactive epitope was detected. Conclusions This is the first report of autoreactive anti-actin antibodies involved in the pathogenesis of autoimmune haemolytic anaemia.

  1. A new F-actin structure in fungi: actin ring formation around the cell nucleus of Cryptococcus neoformans.

    Science.gov (United States)

    Kopecká, Marie; Kawamoto, Susumu; Yamaguchi, Masashi

    2013-04-01

    The F-actin cytoskeleton of Cryptococcus neoformans is known to comprise actin cables, cortical patches and cytokinetic ring. Here, we describe a new F-actin structure in fungi, a perinuclear F-actin collar ring around the cell nucleus, by fluorescent microscopic imaging of rhodamine phalloidin-stained F-actin. Perinuclear F-actin rings form in Cryptococcus neoformans treated with the microtubule inhibitor Nocodazole or with the drug solvent dimethyl sulfoxide (DMSO) or grown in yeast extract peptone dextrose (YEPD) medium, but they are absent in cells treated with Latrunculin A. Perinuclear F-actin rings may function as 'funicular cabin' for the cell nucleus, and actin cables as intracellular 'funicular' suspending nucleus in the central position in the cell and moving nucleus along the polarity axis along actin cables.

  2. Spontaneous actin dynamics in contractile rings

    Science.gov (United States)

    Kruse, Karsten; Wollrab, Viktoria; Thiagarajan, Raghavan; Wald, Anne; Riveline, Daniel

    Networks of polymerizing actin filaments are known to be capable to self-organize into a variety of structures. For example, spontaneous actin polymerization waves have been observed in living cells in a number of circumstances, notably, in crawling neutrophils and slime molds. During later stages of cell division, they can also spontaneously form a contractile ring that will eventually cleave the cell into two daughter cells. We present a framework for describing networks of polymerizing actin filaments, where assembly is regulated by various proteins. It can also include the effects of molecular motors. We show that the molecular processes driven by these proteins can generate various structures that have been observed in contractile rings of fission yeast and mammalian cells. We discuss a possible functional role of each of these patterns. The work was supported by Agence Nationale de la Recherche, France, (ANR-10-LABX-0030-INRT) and by Deutsche Forschungsgemeinschaft through SFB1027.

  3. Superficial tension: experimental model with simple materials

    Directory of Open Access Journals (Sweden)

    Tintori Ferreira, María Alejandra

    2012-09-01

    Full Text Available In this work appears a didactic offer based on an experimental activity using materials of very low cost, orientated to achieving that the student understand and interpret the phenomenon of superficial tension together with the importance of the modeling in sciences. It has as principal aim of education bring the student over to the mechanics of the static fluids and the intermolecular forces, combining scientific contents with questions near to the student what provides an additional motivation to the reflection of the scientific investigation.

  4. Superficial Vein Thrombophlebitis in a Football Athlete.

    Science.gov (United States)

    Schleich, Kevin T; Smoot, M Kyle

    2016-03-01

    A 22-year-old professional football player presented to a preparticipation physical examination with a 2-week history of left leg discomfort extending from the groin to the knee over the previous 2 weeks. He was found to have superficial vein thrombophlebitis (SVT) of the left great saphenous vein extending from the knee to within approximately 1.6 cm of the saphenofemoral junction. There is paucity in the literature regarding the management of SVT, particularly in actively training athletes. This case addresses the considerations of anticoagulation management for SVT as well as the unique challenge of managing anticoagulation therapy in an athlete that is actively training.

  5. Temperature distributions in 136 superficial radiothermotherapies

    International Nuclear Information System (INIS)

    Willich, N.; Duve, S.; Pfluger, T.; Bachmeier, K.

    1992-01-01

    Temperature distributions from 136 superficial radiothermotherapies in patients were analysed and three-dimensionally reconstructed. The calculation of mean values and standard deviations of the temperature measuring probes considering water bolus temperature, master probe temperature, site of the probes relatively to different applicator positions and site of the probes in the heated tissues yielded satisfactory temperature distributions for chest wall treatment in contrast to other regions of the body. Radiothermotherapy was statistically not superior to radiotherapy alone with respect to local tumor control. (authors)

  6. Superficial femoral artery: current treatment options

    International Nuclear Information System (INIS)

    Tepe, Gunnar; Schmehl, Joerg; Heller, Stephan; Wiesinger, Benjamin; Claussen, Claus D.; Duda, Stephan H.

    2006-01-01

    Treatment of the superficial femoral artery (SFA) has been among the least effective of all endovascular procedures in terms of long-term patency. The relatively small vessel lumen, in conjunction with a high plaque burden, slow flow, and a high frequency of primary occlusions, contributes to a considerable rate of acute technical failures. Because of these technical limitations a much effort has been made during the past years. This manuscript should summarize the hopes and limitations of different approaches such as brachytherapy, cutting balloons, stents and stent grafts, drug-eluting stents, and drug-coated balloons. (orig.)

  7. The Bacterial Actin MamK

    Science.gov (United States)

    Ozyamak, Ertan; Kollman, Justin; Agard, David A.; Komeili, Arash

    2013-01-01

    It is now recognized that actin-like proteins are widespread in bacteria and, in contrast to eukaryotic actins, are highly diverse in sequence and function. The bacterial actin, MamK, represents a clade, primarily found in magnetotactic bacteria, that is involved in the proper organization of subcellular organelles, termed magnetosomes. We have previously shown that MamK from Magnetospirillum magneticum AMB-1 (AMB-1) forms dynamic filaments in vivo. To gain further insights into the molecular mechanisms that underlie MamK dynamics and function, we have now studied the in vitro properties of MamK. We demonstrate that MamK is an ATPase that, in the presence of ATP, assembles rapidly into filaments that disassemble once ATP is depleted. The mutation of a conserved active site residue (E143A) abolishes ATPase activity of MamK but not its ability to form filaments. Filament disassembly depends on both ATPase activity and potassium levels, the latter of which results in the organization of MamK filaments into bundles. These data are consistent with observations indicating that accessory factors are required to promote filament disassembly and for spatial organization of filaments in vivo. We also used cryo-electron microscopy to obtain a high resolution structure of MamK filaments. MamK adopts a two-stranded helical filament architecture, but unlike eukaryotic actin and other actin-like filaments, subunits in MamK strands are unstaggered giving rise to a unique filament architecture. Beyond extending our knowledge of the properties and function of MamK in magnetotactic bacteria, this study emphasizes the functional and structural diversity of bacterial actins in general. PMID:23204522

  8. HIV infection of T cells: actin-in and actin-out.

    Science.gov (United States)

    Liu, Yin; Belkina, Natalya V; Shaw, Stephen

    2009-04-14

    Three studies shed light on the decade-old observation that the actin cytoskeleton is hijacked to facilitate entry of HIV into its target cells. Polymerization of actin is required to assemble high concentrations of CD4 and CXCR4 at the plasma membrane, which promote viral binding and entry in both the simple model of infection by free virus and the more physiologically relevant route of infection through the virological synapse. Three types of actin-interacting proteins-filamin, ezrin/radixin/moesin (ERM), and cofilin-are now shown to play critical roles in this process. Filamin binds to both CD4 and CXCR4 in a manner promoted by signaling of the HIV gp120 glycoprotein. ERM proteins attach actin filaments to the membrane and may promote polymerization of actin. Early in the process of viral entry, cofilin is inactivated, which is proposed to facilitate the early assembly of actin filaments, but cofilin is reported to be activated soon thereafter to facilitate postentry events. This complex role of cofilin may help to reconcile the paradox that actin polymerization promotes initial binding and fusion steps but inhibits some subsequent early postentry events.

  9. Non-Straub type actin from molluscan catch muscle

    Energy Technology Data Exchange (ETDEWEB)

    Shelud' ko, Nikolay S., E-mail: sheludko@stl.ru; Girich, Ulyana V.; Lazarev, Stanislav S.; Vyatchin, Ilya G.

    2016-05-27

    We have developed a method of obtaining natural actin from smooth muscles of the bivalves on the example of the Crenomytilus grayanus catch muscle. The muscles were previously rigorized to prevent a loss of thin filaments during homogenization and washings. Thin filaments were isolated with a low ionic strength solution in the presence of ATP and sodium pyrophosphate. Surface proteins of thin filaments-tropomyosin, troponin, calponin and some minor actin-binding proteins-were dissociated from actin filaments by increasing the ionic strength to 0.6 M KCL. Natural fibrillar actin obtained in that way depolymerizes easily in low ionic strength solutions commonly used for the extraction of Straub-type actin from acetone powder. Purification of natural actin was carried out by the polymerization–depolymerization cycle. The content of inactivated actin remaining in the supernatant is much less than at a similar purification of Straub-type actin. A comparative investigation was performed between the natural mussel actin and the Straub-type rabbit skeletal actin in terms of the key properties of actin: polymerization, activation of Mg-ATPase activity of myosin, and the electron-microscopic structure of actin polymers. -- Highlights: •We developed method of repolymerizable invertebrate smooth muscle actin obtaining. •Our method does not involve use of denaturating agents, which could modify proteins. •Viscosity and polymerization rate of actin, gained that way, is similar to Straub one. •Electron microscopy showed that repolymerized mussel actin is similar to Straub one. •Repolymerized mussel actin has greater ATPase activating capacity, than Straub actin.

  10. Superficial basal cell carcinoma: A comparison of superficial only subtype with superficial combined with other subtypes by age, sex and anatomic site in 3150 cases.

    Science.gov (United States)

    Pyne, John H; Myint, Esther; Barr, Elizabeth M; Clark, Simon P; David, Michael; Na, Renua; Hou, Ruihang

    2017-08-01

    Basal cell carcinoma (BCC) may present as superficial subtype alone (sBCC) or superficial combined with other subtypes. The objective of this study was to compare sBCC without or with other BCC subtypes by age, sex and anatomic site. We retrospectively collected superficial BCC with the above characteristics from an Australian center during 2009 to 2014. We recorded 1528 sBCC and 1622 superficial BCC combined with other BCC subtype cases. Males numbered 2007 and females 1140. On males, head sites (forehead, cheek, nose and ear combined) compared to limb plus trunk sites displayed a higher incidence of superficial BCC combined with either nodular and or aggressive BCC subtypes (OR 13.15 CI 95% 8.9-19.5 P < .0001). On females a similar comparison also found a higher incidence of superficial BCC combined with solid subtype BCC on head sites compared to trunk and limb sites (OR 9.66 CI 95% 5.8-16.1 P < .0001). Superficial BCC alone is more likely on younger females on trunk and limb sites. Small partial biopsies reported as sBCC may miss other BCC subtypes present with higher risk on facial sites for males and females. Males had smaller proportions of superficial only subtype BCC on facial and ear sites compared to females. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  11. Actin dynamics and the elasticity of cytoskeletal networks

    Directory of Open Access Journals (Sweden)

    2009-09-01

    Full Text Available The structural integrity of a cell depends on its cytoskeleton, which includes an actin network. This network is transient and depends upon the continual polymerization and depolymerization of actin. The degradation of an actin network, and a corresponding reduction in cell stiffness, can indicate the presence of disease. Numerical simulations will be invaluable for understanding the physics of these systems and the correlation between actin dynamics and elasticity. Here we develop a model that is capable of generating actin network structures. In particular, we develop a model of actin dynamics which considers the polymerization, depolymerization, nucleation, severing, and capping of actin filaments. The structures obtained are then fed directly into a mechanical model. This allows us to qualitatively assess the effects of changing various parameters associated with actin dynamics on the elasticity of the material.

  12. Integrins in cell migration – the actin connection

    OpenAIRE

    Vicente-Manzanares, Miguel; Choi, Colin Kiwon; Horwitz, Alan Rick

    2008-01-01

    The connection between integrins and actin is driving the field of cell migration in new directions. Integrins and actin are coupled through a physical linkage, which provides traction for migration. Recent studies show the importance of this linkage in regulating adhesion organization and development. Actin polymerization orchestrates adhesion assembly near the leading edge of a migrating cell, and the dynamic cross-linking of actin filaments promotes adhesion maturat...

  13. Curvature and torsion in growing actin networks

    International Nuclear Information System (INIS)

    Shaevitz, Joshua W; Fletcher, Daniel A

    2008-01-01

    Intracellular pathogens such as Listeria monocytogenes and Rickettsia rickettsii move within a host cell by polymerizing a comet-tail of actin fibers that ultimately pushes the cell forward. This dense network of cross-linked actin polymers typically exhibits a striking curvature that causes bacteria to move in gently looping paths. Theoretically, tail curvature has been linked to details of motility by considering force and torque balances from a finite number of polymerizing filaments. Here we track beads coated with a prokaryotic activator of actin polymerization in three dimensions to directly quantify the curvature and torsion of bead motility paths. We find that bead paths are more likely to have low rather than high curvature at any given time. Furthermore, path curvature changes very slowly in time, with an autocorrelation decay time of 200 s. Paths with a small radius of curvature, therefore, remain so for an extended period resulting in loops when confined to two dimensions. When allowed to explore a three-dimensional (3D) space, path loops are less evident. Finally, we quantify the torsion in the bead paths and show that beads do not exhibit a significant left- or right-handed bias to their motion in 3D. These results suggest that paths of actin-propelled objects may be attributed to slow changes in curvature, possibly associated with filament debranching, rather than a fixed torque

  14. The nature of the globular- to fibrous-actin transition.

    Science.gov (United States)

    Oda, Toshiro; Iwasa, Mitsusada; Aihara, Tomoki; Maéda, Yuichiro; Narita, Akihiro

    2009-01-22

    Actin plays crucial parts in cell motility through a dynamic process driven by polymerization and depolymerization, that is, the globular (G) to fibrous (F) actin transition. Although our knowledge about the actin-based cellular functions and the molecules that regulate the G- to F-actin transition is growing, the structural aspects of the transition remain enigmatic. We created a model of F-actin using X-ray fibre diffraction intensities obtained from well oriented sols of rabbit skeletal muscle F-actin to 3.3 A in the radial direction and 5.6 A along the equator. Here we show that the G- to F-actin conformational transition is a simple relative rotation of the two major domains by about 20 degrees. As a result of the domain rotation, the actin molecule in the filament is flat. The flat form is essential for the formation of stable, helical F-actin. Our F-actin structure model provides the basis for understanding actin polymerization as well as its molecular interactions with actin-binding proteins.

  15. Changes in Actin Organization During the Cytotoxic Process

    NARCIS (Netherlands)

    Radosevic, K.; Radosevic, Katarina; van Leeuwen, Anne Marie T.; Segers-Nolten, Gezina M.J.; Figdor, Carl; de Grooth, B.G.; Greve, Jan

    1994-01-01

    Changes in organization of F-actin during the cytotoxic process between NK and K562 cells have been observed and studied using confpcal laser scanning microscopy and quantitative fluorescence microscopy. An increase in F-actin content and orientation of F-actin towards the target cell have been

  16. Adhesive F-actin Waves: A Novel Integrin-Mediated Adhesion Complex Coupled to Ventral Actin Polymerization

    OpenAIRE

    Case, Lindsay B.; Waterman, Clare M.

    2011-01-01

    At the leading lamellipodium of migrating cells, protrusion of an Arp2/3-nucleated actin network is coupled to formation of integrin-based adhesions, suggesting that Arp2/3-mediated actin polymerization and integrin-dependent adhesion may be mechanistically linked. Arp2/3 also mediates actin polymerization in structures distinct from the lamellipodium, in "ventral F-actin waves" that propagate as spots and wavefronts along the ventral plasma membrane. Here we show that integrins engage the ex...

  17. Tailor-made ezrin actin binding domain to probe its interaction with actin in-vitro.

    Directory of Open Access Journals (Sweden)

    Rohini Shrivastava

    Full Text Available Ezrin, a member of the ERM (Ezrin/Radixin/Moesin protein family, is an Actin-plasma membrane linker protein mediating cellular integrity and function. In-vivo study of such interactions is a complex task due to the presence of a large number of endogenous binding partners for both Ezrin and Actin. Further, C-terminal actin binding capacity of the full length Ezrin is naturally shielded by its N-terminal, and only rendered active in the presence of Phosphatidylinositol bisphosphate (PIP2 or phosphorylation at the C-terminal threonine. Here, we demonstrate a strategy for the design, expression and purification of constructs, combining the Ezrin C-terminal actin binding domain, with functional elements such as fusion tags and fluorescence tags to facilitate purification and fluorescence microscopy based studies. For the first time, internal His tag was employed for purification of Ezrin actin binding domain based on in-silico modeling. The functionality (Ezrin-actin interaction of these constructs was successfully demonstrated by using Total Internal Reflection Fluorescence Microscopy. This design can be extended to other members of the ERM family as well.

  18. Social network extraction based on Web: 1. Related superficial methods

    Science.gov (United States)

    Khairuddin Matyuso Nasution, Mahyuddin

    2018-01-01

    Often the nature of something affects methods to resolve the related issues about it. Likewise, methods to extract social networks from the Web, but involve the structured data types differently. This paper reveals several methods of social network extraction from the same sources that is Web: the basic superficial method, the underlying superficial method, the description superficial method, and the related superficial methods. In complexity we derive the inequalities between methods and so are their computations. In this case, we find that different results from the same tools make the difference from the more complex to the simpler: Extraction of social network by involving co-occurrence is more complex than using occurrences.

  19. Antibody conjugate radioimmunotherapy of superficial bladder cancer

    International Nuclear Information System (INIS)

    Perkins, Alan; Hopper, Melanie; Murray, Andrea; Frier, Malcolm; Bishop, Mike

    2002-01-01

    The administration of antibody conjugates for cancer therapy is now proving to be of clinical value. We are currently undertaking a programme of clinical studies using the monoclonal antibody C 595 (gG3) which reacts with the MUC1 glycoprotein antigen that is aberrantly expressed in a high proportion of bladder tumours. Radio immuno conjugates of the C 595 antibody have been produced with high radiolabelling efficiency and immuno reactivity using Tc-99 m and In-111 for diagnostic imaging, and disease staging and the cytotoxic radionuclides Cu-67 and Re-188 for therapy of superficial bladder cancer. A Phase I/II therapeutic trail involving the intravesical administration of antibody directly into the bladder has now begun. (author)

  20. Superficial biopsy of the cervix: new technique

    Directory of Open Access Journals (Sweden)

    Ettore De Girolami

    2002-06-01

    Full Text Available Cytological examination of the cervix in all patients is considered a necessity. A new technique of superficial biopsy is described and advantages en. numeratedEl Autor presenta una nueva técnica para biopsia superficial, considerando la necesidad de hacer rutinariamente a todas las pacientes el examen citológico del cérvix. La muestra se toma con el extremo de un aplicador cubierto de gaza. Este dispositivo, humedecido previamente en una solución de etanol, metanol y éter etílico, se aplica inmediatamente al cérvix del útero y, dándole un movimiento como el de las manecillas del reloj, se toma la muestra con la que se hacen frotis por aposición que se colorean por el método de Giemsa. Esta técnica ofrece la ventaja de que el tejido en estudio se adhiere perfectamente a la gaza, la cual, con el movimiento de rotación, desprende las células del "os uteri", las que son fijadas al mismo tiempo. La preparación del dispositivo es sumamente sencilla. La prueba resulta de bajo costo como de breve y fácil realizaciónL' Autore prende in considerazione l'utilitá dell' esame citologico del collo dell'utero, fatto rutinariamente a tutte le pazienti. Si descrive una nuova tecnica di biopsia superficiale che oltfe ad offrire dei vantaggi é di poca spesa e si puó eseguire con facilitá ed in breve tempo

  1. Volumetric and superficial characterization of carbon activated; Caracterizacion volumetrica y superficial de carbon activado

    Energy Technology Data Exchange (ETDEWEB)

    Carrera G, L.M.; Garcia S, I.; Jimenez B, J.; Solache R, M.; Lopez M, B.; Bulbulian G, S.; Olguin G, M.T. [Departamento de Quimica, Gerencia de Ciencias Basicas, Instituto Nacional de Investigaciones Nucleares, A.P. 18-1027, 11801 Mexico D.F. (Mexico)

    2000-07-01

    The activated carbon is the resultant material of the calcination process of natural carbonated materials as coconut shells or olive little bones. It is an excellent adsorbent of diluted substances, so much in colloidal form, as in particles form. Those substances are attracted and retained by the carbon surface. In this work is make the volumetric and superficial characterization of activated carbon treated thermically (300 Centigrade) in function of the grain size average. (Author)

  2. Cytoplasmic Actin: Purification and Single Molecule Assembly Assays

    Science.gov (United States)

    Hansen, Scott D.; Zuchero, J. Bradley; Mullins, R. Dyche

    2014-01-01

    The actin cytoskeleton is essential to all eukaryotic cells. In addition to playing important structural roles, assembly of actin into filaments powers diverse cellular processes, including cell motility, cytokinesis, and endocytosis. Actin polymerization is tightly regulated by its numerous cofactors, which control spatial and temporal assembly of actin as well as the physical properties of these filaments. Development of an in vitro model of actin polymerization from purified components has allowed for great advances in determining the effects of these proteins on the actin cytoskeleton. Here we describe how to use the pyrene actin assembly assay to determine the effect of a protein on the kinetics of actin assembly, either directly or as mediated by proteins such as nucleation or capping factors. Secondly, we show how fluorescently labeled phalloidin can be used to visualize the filaments that are created in vitro to give insight into how proteins regulate actin filament structure. Finally, we describe a method for visualizing dynamic assembly and disassembly of single actin filaments and fluorescently labeled actin binding proteins using total internal reflection fluorescence (TIRF) microscopy. PMID:23868587

  3. Bundling Actin Filaments From Membranes: Some Novel Players

    Directory of Open Access Journals (Sweden)

    Clément eThomas

    2012-08-01

    Full Text Available Progress in live-cell imaging of the cytoskeleton has significantly extended our knowledge about the organization and dynamics of actin filaments near the plasma membrane of plant cells. Noticeably, two populations of filamentous structures can be distinguished. On the one hand, fine actin filaments which exhibit an extremely dynamic behavior basically characterized by fast polymerization and prolific severing events, a process referred to as actin stochastic dynamics. On the other hand, thick actin bundles which are composed of several filaments and which are comparatively more stable although they constantly remodel as well. There is evidence that the actin cytoskeleton plays critical roles in trafficking and signaling at both the cell cortex and organelle periphery but the exact contribution of actin bundles remains unclear. A common view is that actin bundles provide the long-distance tracks used by myosin motors to deliver their cargo to growing regions and accordingly play a particularly important role in cell polarization. However, several studies support that actin bundles are more than simple passive highways and display multiple and dynamic roles in the regulation of many processes, such as cell elongation, polar auxin transport, stomatal and chloroplast movement, and defense against pathogens. The list of identified plant actin-bundling proteins is ever expanding, supporting that plant cells shape structurally and functionally different actin bundles. Here I review the most recently characterized actin-bundling proteins, with a particular focus on those potentially relevant to membrane trafficking and/or signaling.

  4. Actin-interacting Protein 1 Promotes Disassembly of Actin-depolymerizing Factor/Cofilin-bound Actin Filaments in a pH-dependent Manner.

    Science.gov (United States)

    Nomura, Kazumi; Hayakawa, Kimihide; Tatsumi, Hitoshi; Ono, Shoichiro

    2016-03-04

    Actin-interacting protein 1 (AIP1) is a conserved WD repeat protein that promotes disassembly of actin filaments when actin-depolymerizing factor (ADF)/cofilin is present. Although AIP1 is known to be essential for a number of cellular events involving dynamic rearrangement of the actin cytoskeleton, the regulatory mechanism of the function of AIP1 is unknown. In this study, we report that two AIP1 isoforms from the nematode Caenorhabditis elegans, known as UNC-78 and AIPL-1, are pH-sensitive in enhancement of actin filament disassembly. Both AIP1 isoforms only weakly enhance disassembly of ADF/cofilin-bound actin filaments at an acidic pH but show stronger disassembly activity at neutral and basic pH values. However, a severing-defective mutant of UNC-78 shows pH-insensitive binding to ADF/cofilin-decorated actin filaments, suggesting that the process of filament severing or disassembly, but not filament binding, is pH-dependent. His-60 of AIP1 is located near the predicted binding surface for the ADF/cofilin-actin complex, and an H60K mutation of AIP1 partially impairs its pH sensitivity, suggesting that His-60 is involved in the pH sensor for AIP1. These biochemical results suggest that pH-dependent changes in AIP1 activity might be a novel regulatory mechanism of actin filament dynamics. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  5. Wnt Signalling Promotes Actin Dynamics during Axon Remodelling through the Actin-Binding Protein Eps8.

    Directory of Open Access Journals (Sweden)

    Eleanna Stamatakou

    Full Text Available Upon arrival at their synaptic targets, axons slow down their growth and extensively remodel before the assembly of presynaptic boutons. Wnt proteins are target-derived secreted factors that promote axonal remodelling and synaptic assembly. In the developing spinal cord, Wnts secreted by motor neurons promote axonal remodelling of NT-3 responsive dorsal root ganglia neurons. Axon remodelling induced by Wnts is characterised by growth cone pausing and enlargement, processes that depend on the re-organisation of microtubules. However, the contribution of the actin cytoskeleton has remained unexplored. Here, we demonstrate that Wnt3a regulates the actin cytoskeleton by rapidly inducing F-actin accumulation in growth cones from rodent DRG neurons through the scaffold protein Dishevelled-1 (Dvl1 and the serine-threonine kinase Gsk3β. Importantly, these changes in actin cytoskeleton occurs before enlargement of the growth cones is evident. Time-lapse imaging shows that Wnt3a increases lamellar protrusion and filopodia velocity. In addition, pharmacological inhibition of actin assembly demonstrates that Wnt3a increases actin dynamics. Through a yeast-two hybrid screen, we identified the actin-binding protein Eps8 as a direct interactor of Dvl1, a scaffold protein crucial for the Wnt signalling pathway. Gain of function of Eps8 mimics Wnt-mediated axon remodelling, whereas Eps8 silencing blocks the axon remodelling activity of Wnt3a. Importantly, blockade of the Dvl1-Eps8 interaction completely abolishes Wnt3a-mediated axonal remodelling. These findings demonstrate a novel role for Wnt-Dvl1 signalling through Eps8 in the regulation of axonal remodeling.

  6. Distinct functional interactions between actin isoforms and nonsarcomeric myosins.

    Directory of Open Access Journals (Sweden)

    Mirco Müller

    Full Text Available Despite their near sequence identity, actin isoforms cannot completely replace each other in vivo and show marked differences in their tissue-specific and subcellular localization. Little is known about isoform-specific differences in their interactions with myosin motors and other actin-binding proteins. Mammalian cytoplasmic β- and γ-actin interact with nonsarcomeric conventional myosins such as the members of the nonmuscle myosin-2 family and myosin-7A. These interactions support a wide range of cellular processes including cytokinesis, maintenance of cell polarity, cell adhesion, migration, and mechano-electrical transduction. To elucidate differences in the ability of isoactins to bind and stimulate the enzymatic activity of individual myosin isoforms, we characterized the interactions of human skeletal muscle α-actin, cytoplasmic β-actin, and cytoplasmic γ-actin with human myosin-7A and nonmuscle myosins-2A, -2B and -2C1. In the case of nonmuscle myosins-2A and -2B, the interaction with either cytoplasmic actin isoform results in 4-fold greater stimulation of myosin ATPase activity than was observed in the presence of α-skeletal muscle actin. Nonmuscle myosin-2C1 is most potently activated by β-actin and myosin-7A by γ-actin. Our results indicate that β- and γ-actin isoforms contribute to the modulation of nonmuscle myosin-2 and myosin-7A activity and thereby to the spatial and temporal regulation of cytoskeletal dynamics. FRET-based analyses show efficient copolymerization abilities for the actin isoforms in vitro. Experiments with hybrid actin filaments show that the extent of actomyosin coupling efficiency can be regulated by the isoform composition of actin filaments.

  7. Profilin connects actin assembly with microtubule dynamics

    Czech Academy of Sciences Publication Activity Database

    Nejedla, M.; Sadi, S.; Sulimenko, Vadym; de Almeida, F.N.; Blom, H.; Dráber, Pavel; Aspenstrom, P.; Karlsson, R.

    2016-01-01

    Roč. 27, č. 15 (2016), s. 2381-2393 ISSN 1059-1524 R&D Projects: GA ČR GA16-25159S Institutional support: RVO:68378050 Keywords : cross-linked profilin * arp2/3 complex * f-actin * microfilament system * migrating cells * focal adhesions * cultured-cells * messenger-rna * living cells * protein Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.685, year: 2016

  8. Computational Study of the Binding Mechanism of Actin-Depolymerizing Factor 1 with Actin in Arabidopsis thaliana.

    Directory of Open Access Journals (Sweden)

    Juan Du

    Full Text Available Actin is a highly conserved protein. It plays important roles in cellular function and exists either in the monomeric (G-actin or polymeric form (F-actin. Members of the actin-depolymerizing factor (ADF/cofilin protein family bind to both G-actin and F-actin and play vital roles in actin dynamics by manipulating the rates of filament polymerization and depolymerization. It has been reported that the S6D and R98A/K100A mutants of actin-depolymerizing factor 1 (ADF1 in Arabidopsis thaliana decreased the binding affinity of ADF for the actin monomer. To investigate the binding mechanism and dynamic behavior of the ADF1-actin complex, we constructed a homology model of the AtADF1-actin complex based on the crystal structure of AtADF1 and the twinfilin C-terminal ADF-H domain in a complex with a mouse actin monomer. The model was then refined for subsequent molecular dynamics simulations. Increased binding energy of the mutated system was observed using the Molecular Mechanics Generalized Born Surface Area and Poisson-Boltzmann Surface Area (MM-GB/PBSA methods. To determine the residues that make decisive contributions to the ADF1 actin-binding affinity, per-residue decomposition and computational alanine scanning analyses were performed, which provided more detailed information on the binding mechanism. Root-mean-square fluctuation and principal component analyses confirmed that the S6D and R98A/K100A mutants induced an increased conformational flexibility. The comprehensive molecular insight gained from this study is of great importance for understanding the binding mechanism of ADF1 and G-actin.

  9. Size exclusion chromatography with superficially porous particles.

    Science.gov (United States)

    Schure, Mark R; Moran, Robert E

    2017-01-13

    A comparison is made using size-exclusion chromatography (SEC) of synthetic polymers between fully porous particles (FPPs) and superficially porous particles (SPPs) with similar particle diameters, pore sizes and equal flow rates. Polystyrene molecular weight standards with a mobile phase of tetrahydrofuran are utilized for all measurements conducted with standard HPLC equipment. Although it is traditionally thought that larger pore volume is thermodynamically advantageous in SEC for better separations, SPPs have kinetic advantages and these will be shown to compensate for the loss in pore volume compared to FPPs. The comparison metrics include the elution range (smaller with SPPs), the plate count (larger for SPPs), the rate production of theoretical plates (larger for SPPs) and the specific resolution (larger with FPPs). Advantages to using SPPs for SEC are discussed such that similar separations can be conducted faster using SPPs. SEC using SPPs offers similar peak capacities to that using FPPs but with faster operation. This also suggests that SEC conducted in the second dimension of a two-dimensional liquid chromatograph may benefit with reduced run time and with equivalently reduced peak width making SPPs advantageous for sampling the first dimension by the second dimension separator. Additional advantages are discussed for biomolecules along with a discussion of optimization criteria for size-based separations. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Superficial siderosis: Case report and literature review

    Directory of Open Access Journals (Sweden)

    Kresojević Nikola D.

    2013-01-01

    Full Text Available Introduction. Superficial siderosis (SS is caused by chronic subarachnoid bleeding and is characterized by free iron and hemosiderin deposition along the pial and subpial structures of central nervous system. SS leads to progressive and irreversible CNS damage. The most common causes of chronic subarachnoidal bleeding are tumors, head and spinal cord trauma, arteriovenous malformations and aneurysms. SS is characterized by clinical triad: sensorineural hearing loss, cerebellar ataxia and piramydal signs. Brain MR imaging is the investigation of choice for the diagnosis of SS. Typical findings include hypointensities seen on T2­weighted MR imaging around the brain, cerebellum, brain stem, spinal cord, VIII cranial nerve and atrophy of cerebellum and medulla. Case Outline. A 71-year­old female patient noticed hand tremor in the middle of the third decade of life, and later slowly progressive bilateral hearing loss. At the age of 64 she developed un­ steady gate, hand clumsiness and dysarthria, to became incapable of independent walking and standing five years later. Clinical course and brain MRI findings were typical for SS, but additional investigation did not reveal the couse of subarahnoidal bleeding. Conclusion. SS represents a rare and under­recognized condition that must be considered in all patients with cerebellar syndrome of unknown cause. Early diagnosis of SS in some cases with identified cause of chronic bleeding allowes therapeutic interventions that may prevent further progression of the disease. [Projekat Ministarstva nauke Republike Srbije, br. 175090

  11. Modified Functional Superficial Parotidectomy With Ligation of the Major Branch of the Parotid Duct Extending to the Superficial Lobe.

    Science.gov (United States)

    Chang, Jung Woo; Leem, Soo Seong; Choi, Hwan Jun; Lee, Jang Hyun

    2017-05-01

    A functional superficial parotidectomy can maintain salivary function by preserving the Stensen duct. However, this technique still brings the possibility of salivary leakage, because major branches of the parotid duct from the resected site do not get ligated. To reduce this complication, this study introduces a modified technique with major branch ligation. From December 2008 to February 2015, 14 patients who underwent superficial parotidectomy were divided into 2 groups. Group A was treated with the modified functional superficial parotidectomy involving the major branch between the superficial lobe and parotid duct. Group B was treated with the conventional superficial parotidectomy without involving the major branch of the parotid duct. The clinical complications, period of Hemovac usage, and surgical duration were noted in each group. Two of 8 patients in group A had a major branch from Stensen duct that was ligated, and there was no evidence of salivary leakage or sialocele in any of the patients of group A, whereas group B contained 2 cases of salivary leakage, one of which became sialocele. Group A had a significantly longer Hemovac maintenance period than group B (P < 0.05), and the duration of surgery was also significantly different between the 2 groups (P < 0.05). Because a solitary major branch of the main parotid duct occasionally extends toward the superficial lobe, our modified technique-functional superficial parotidectomy with ligation of the major branch toward the superficial lobe-is a useful option for treatment of a benign parotid mass in such cases.

  12. Fibroblast-mediated contraction in actinically exposed and actinically protected aging skin

    International Nuclear Information System (INIS)

    Marks, M.W.; Morykwas, M.J.; Wheatley, M.J.

    1990-01-01

    The changes in skin morphology over time are a consequence of both chronologic aging and the accumulation of environmental exposure. Through observation, we know that actinic radiation intensifies the apparent aging of skin. We have investigated the effects of aging and actinic radiation on the ability of fibroblasts to contract collagen-fibroblast lattices. Preauricular and postauricular skin samples were obtained from eight patients aged 49 to 74 undergoing rhytidectomy. The samples were kept separate, and the fibroblasts were grown in culture. Lattices constructed with preauricular fibroblasts consistently contracted more than lattices containing postauricular fibroblasts. The difference in amount of contraction in 7 days between sites was greatest for the younger patients and decreased linearly as donor age increased (r = -0.96). This difference may be due to preauricular fibroblasts losing their ability to contract a lattice as aging skin is exposed to more actinic radiation

  13. Characterization of a porcine model of chronic superficial varicose veins.

    Science.gov (United States)

    Jones, Gregory T; Grant, Mark W; Thomson, Ian A; Hill, B Geraldine; van Rij, André M

    2009-06-01

    Previous animal models of venous disease, while inducing venous hypertension and valvular insufficiency, do not produce superficial varicose veins. In this study, we aimed to develop and characterize a pig-based model of superficial varicose veins. Right femoral arteriovenous fistulae (AVF) were surgically fashioned in young adult pigs. Animals were examined at postoperative times up to 15 weeks to determine the development of varicose veins and measurement of both blood pressure and flow velocities within the superficial thigh veins. Histology and vascular corrosion casts were used to characterize the resulting structural venous alterations. Porcine pathophysiological features were compared with those of human primary superficial varicose veins. Gross superficial varicosities developed over the ipsilateral medial thigh region after an initial lag period of 1-2 weeks. Veins demonstrated retrograde filling with valvular incompetence, and a moderate, non-pulsatile, venous hypertension, which was altered by changes in posture and Valsalva. Venous blood flow velocities were elevated to 15-30 cm/s in varicose veins. Structurally, pig varicose veins were enlarged, tortuous, had valvular degeneration, and regions of focal medial atrophy with or without overlying intimal thickening. The superficial varicose veins, which developed within this model, have a pathophysiology that is consistent with that observed in humans. The porcine femoral AVF model is proposed as a suitable experimental model to evaluate the pathobiology of superficial venous disease. It may also be suitable for the evaluation of treatment interventions including drug therapy.

  14. Hemipelvic irradiation for superficial bladder cancer

    International Nuclear Information System (INIS)

    Tashiro, Kazuya; Machida, Toyohei; Ooishi, Yukihiko; Ueda, Masataka; Kido, Akira; Wada, Tetsuro; Yoshigoe, Fukuo; Yamashita, Takashi; Mochizuki, Sachio

    1985-01-01

    In 15 patients with superficial bladder cancer hemipelvic irradiation was performed for prevention of relapse of cancer and decrease in side effects with following results. All patients received TUR-Bt at our department during the six years period from 1978 to 1983. As to stages, one was classified as Ta, 11 as T 1 and 3 as T 2, and pathologic diagnosis was transitional epithelial carcinoma of grade 1 in 1 case, grade 2 in 8 cases and grade 3 in 6 cases. Irradiation was started from the 7 th to 14 th day after TUR-Bt. At first, hemipelvic anterior and posterior field including the field from the affected pelvis to 1 to 2 cm beyond the midline toward the contralateral side and from the aortic bifurcation to the prostatic urethra were irradiated at a dose of 45 Gy. Then, whole bladder field was given an additional rotation irradiation of 20 Gy. The mean observation period was 43 months (ranging from 12 to 79 months) and relapse of cancer was observed in 6 cases out of 15 cases (40%). The site of relapse was in the irradiated site in 2 cases, contralateral site in 3 cases and both side in 1 cases. However, in all of the relapsed cases no aggravation in differential degree or progression in stage was observed. As the side effects, radiation cystitis developed as a delayed damage in 1 case. Thus, although no efficacy for prevention of relapse which we had expected was not seen, this irradiation method effectively inhibited the progression of lesion and development of delayed damage. (author)

  15. Hemipelvic irradiation for superficial bladder cancer

    Energy Technology Data Exchange (ETDEWEB)

    Tashiro, Kazuya; Machida, Toyohei; Ooishi, Yukihiko; Ueda, Masataka; Kido, Akira; Wada, Tetsuro; Yoshigoe, Fukuo; Yamashita, Takashi; Mochizuki, Sachio

    1985-02-01

    In 15 patients with superficial bladder cancer hemipelvic irradiation was performed for prevention of relapse of cancer and decrease in side effects with following results. All patients received TUR-Bt at our department during the six years period from 1978 to 1983. As to stages, one was classified as Ta, 11 as T 1 and 3 as T 2, and pathologic diagnosis was transitional epithelial carcinoma of grade 1 in 1 case, grade 2 in 8 cases and grade 3 in 6 cases. Irradiation was started from the 7 th to 14 th day after TUR-Bt. At first, hemipelvic anterior and posterior field including the field from the affected pelvis to 1 to 2 cm beyond the midline toward the contralateral side and from the aortic bifurcation to the prostatic urethra were irradiated at a dose of 45 Gy. Then, whole bladder field was given an additional rotation irradiation of 20 Gy. The mean observation period was 43 months (ranging from 12 to 79 months) and relapse of cancer was observed in 6 cases out of 15 cases (40%). The site of relapse was in the irradiated site in 2 cases, contralateral site in 3 cases and both side in 1 cases. However, in all of the relapsed cases no aggravation in differential degree or progression in stage was observed. As the side effects, radiation cystitis developed as a delayed damage in 1 case. Thus, although no efficacy for prevention of relapse which we had expected was not seen, this irradiation method effectively inhibited the progression of lesion and development of delayed damage. (author).

  16. Bacterial subversion of host actin dynamics at the plasma membrane.

    Science.gov (United States)

    Carabeo, Rey

    2011-10-01

    Invasion of non-phagocytic cells by a number of bacterial pathogens involves the subversion of the actin cytoskeletal remodelling machinery to produce actin-rich cell surface projections designed to engulf the bacteria. The signalling that occurs to induce these actin-rich structures has considerable overlap among a diverse group of bacteria. The molecular organization within these structures act in concert to internalize the invading pathogen. This dynamic process could be subdivided into three acts - actin recruitment, engulfment, and finally, actin disassembly/internalization. This review will present the current state of knowledge of the molecular processes involved in each stage of bacterial invasion, and provide a perspective that highlights the temporal and spatial control of actin remodelling that occurs during bacterial invasion. © 2011 Blackwell Publishing Ltd.

  17. Stress generation by myosin minifilaments in actin bundles

    International Nuclear Information System (INIS)

    Dasanayake, Nilushi L; Carlsson, Anders E

    2013-01-01

    Forces and stresses generated by the action of myosin minifilaments are analyzed in idealized computer-generated actin bundles, and compared to results for isotropic actin networks. The bundles are generated as random collections of actin filaments in two dimensions with constrained orientations, crosslinked and attached to two fixed walls. Myosin minifilaments are placed on actin filament pairs and allowed to move and deform the network so that it exerts forces on the walls. The vast majority of simulation runs end with contractile minifilament stress, because minifilaments rotate into energetically stable contractile configurations. This process is aided by the bending and stretching of actin filaments, which accomodate minifilament rotation. Stresses for bundles are greater than those for isotropic networks, and antiparallel filaments generate more tension than parallel filaments. The forces transmitted by the actin network to the walls of the simulation cell often exceed the tension in the minifilament itself. (paper)

  18. Measurement and Analysis of in vitro Actin Polymerization

    Science.gov (United States)

    Doolittle, Lynda K.; Rosen, Michael K.; Padrick, Shae B.

    2014-01-01

    Summary The polymerization of actin underlies force generation in numerous cellular processes. While actin polymerization can occur spontaneously, cells maintain control over this important process by preventing actin filament nucleation and then allowing stimulated polymerization and elongation by several regulated factors. Actin polymerization, regulated nucleation and controlled elongation activities can be reconstituted in vitro, and used to probe the signaling cascades cells use to control when and where actin polymerization occurs. Introducing a pyrene fluorophore allows detection of filament formation by an increase in pyrene fluorescence. This method has been used for many years and continues to be broadly used, owing to its simplicity and flexibility. Here we describe how to perform and analyze these in vitro actin polymerization assays, with an emphasis on extracting useful descriptive parameters from kinetic data. PMID:23868594

  19. Coordination of membrane and actin cytoskeleton dynamics during filopodia protrusion.

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    Changsong Yang

    2009-05-01

    Full Text Available Leading edge protrusion of migrating cells involves tightly coordinated changes in the plasma membrane and actin cytoskeleton. It remains unclear whether polymerizing actin filaments push and deform the membrane, or membrane deformation occurs independently and is subsequently stabilized by actin filaments. To address this question, we employed an ability of the membrane-binding I-BAR domain of IRSp53 to uncouple the membrane and actin dynamics and to induce filopodia in expressing cells. Using time-lapse imaging and electron microscopy of IRSp53-I-BAR-expressing B16F1 melanoma cells, we demonstrate that cells are not able to protrude or maintain durable long extensions without actin filaments in their interior, but I-BAR-dependent membrane deformation can create a small and transient space at filopodial tips that is subsequently filled with actin filaments. Moreover, the expressed I-BAR domain forms a submembranous coat that may structurally support these transient actin-free protrusions until they are further stabilized by the actin cytoskeleton. Actin filaments in the I-BAR-induced filopodia, in contrast to normal filopodia, do not have a uniform length, are less abundant, poorly bundled, and display erratic dynamics. Such unconventional structural organization and dynamics of actin in I-BAR-induced filopodia suggests that a typical bundle of parallel actin filaments is not necessary for generation and mechanical support of the highly asymmetric filopodial geometry. Together, our data suggest that actin filaments may not directly drive the protrusion, but only stabilize the space generated by the membrane deformation; yet, such stabilization is necessary for efficient protrusion.

  20. Actin polymerisation at the cytoplasmic face of eukaryotic nuclei

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    David-Watine Brigitte

    2006-05-01

    Full Text Available Abstract Background There exists abundant molecular and ultra-structural evidence to suggest that cytoplasmic actin can physically interact with the nuclear envelope (NE membrane system. However, this interaction has yet to be characterised in living interphase cells. Results Using a fluorescent conjugate of the actin binding drug cytochalasin D (CD-BODIPY we provide evidence that polymerising actin accumulates in vicinity to the NE. In addition, both transiently expressed fluorescent actin and cytoplasmic micro-injection of fluorescent actin resulted in accumulation of actin at the NE-membrane. Consistent with the idea that the cytoplasmic phase of NE-membranes can support this novel pool of perinuclear actin polymerisation we show that isolated, intact, differentiated primary hepatocyte nuclei support actin polymerisation in vitro. Further this phenomenon was inhibited by treatments hindering steric access to outer-nuclear-membrane proteins (e.g. wheat germ agglutinin, anti-nesprin and anti-nucleoporin antibodies. Conclusion We conclude that actin polymerisation occurs around interphase nuclei of living cells at the cytoplasmic phase of NE-membranes.

  1. Electrostatics Control Actin Filament Nucleation and Elongation Kinetics*

    Science.gov (United States)

    Crevenna, Alvaro H.; Naredi-Rainer, Nikolaus; Schönichen, André; Dzubiella, Joachim; Barber, Diane L.; Lamb, Don C.; Wedlich-Söldner, Roland

    2013-01-01

    The actin cytoskeleton is a central mediator of cellular morphogenesis, and rapid actin reorganization drives essential processes such as cell migration and cell division. Whereas several actin-binding proteins are known to be regulated by changes in intracellular pH, detailed information regarding the effect of pH on the actin dynamics itself is still lacking. Here, we combine bulk assays, total internal reflection fluorescence microscopy, fluorescence fluctuation spectroscopy techniques, and theory to comprehensively characterize the effect of pH on actin polymerization. We show that both nucleation and elongation are strongly enhanced at acidic pH, with a maximum close to the pI of actin. Monomer association rates are similarly affected by pH at both ends, although dissociation rates are differentially affected. This indicates that electrostatics control the diffusional encounter but not the dissociation rate, which is critical for the establishment of actin filament asymmetry. A generic model of protein-protein interaction, including electrostatics, explains the observed pH sensitivity as a consequence of charge repulsion. The observed pH effect on actin in vitro agrees with measurements of Listeria propulsion in pH-controlled cells. pH regulation should therefore be considered as a modulator of actin dynamics in a cellular environment. PMID:23486468

  2. Electrostatics control actin filament nucleation and elongation kinetics.

    Science.gov (United States)

    Crevenna, Alvaro H; Naredi-Rainer, Nikolaus; Schönichen, André; Dzubiella, Joachim; Barber, Diane L; Lamb, Don C; Wedlich-Söldner, Roland

    2013-04-26

    The actin cytoskeleton is a central mediator of cellular morphogenesis, and rapid actin reorganization drives essential processes such as cell migration and cell division. Whereas several actin-binding proteins are known to be regulated by changes in intracellular pH, detailed information regarding the effect of pH on the actin dynamics itself is still lacking. Here, we combine bulk assays, total internal reflection fluorescence microscopy, fluorescence fluctuation spectroscopy techniques, and theory to comprehensively characterize the effect of pH on actin polymerization. We show that both nucleation and elongation are strongly enhanced at acidic pH, with a maximum close to the pI of actin. Monomer association rates are similarly affected by pH at both ends, although dissociation rates are differentially affected. This indicates that electrostatics control the diffusional encounter but not the dissociation rate, which is critical for the establishment of actin filament asymmetry. A generic model of protein-protein interaction, including electrostatics, explains the observed pH sensitivity as a consequence of charge repulsion. The observed pH effect on actin in vitro agrees with measurements of Listeria propulsion in pH-controlled cells. pH regulation should therefore be considered as a modulator of actin dynamics in a cellular environment.

  3. Surfing pathogens and the lessons learned for actin polymerization.

    Science.gov (United States)

    Frischknecht, F; Way, M

    2001-01-01

    A number of unrelated bacterial species as well as vaccinia virus (ab)use the process of actin polymerization to facilitate and enhance their infection cycle. Studies into the mechanism by which these pathogens hijack and control the actin cytoskeleton have provided many interesting insights into the regulation of actin polymerization in migrating cells. This review focuses on what we have learnt from the actin-based motilities of Listeria, Shigella and vaccinia and discusses what we would still like to learn from our nasty friends, including enteropathogenic Escherichia coli and Rickettsia

  4. Actin filaments – a target for redox regulation

    Science.gov (United States)

    Wilson, Carlos; Terman, Jonathan R.; González-Billault, Christian; Ahmed, Giasuddin

    2016-01-01

    Actin and its ability to polymerize into dynamic filaments is critical for the form and function of cells throughout the body. While multiple proteins have been characterized as affecting actin dynamics through non-covalent means, actin and its protein regulators are also susceptible to covalent modifications of their amino acid residues. In this regard, oxidation-reduction (Redox) intermediates have emerged as key modulators of the actin cytoskeleton with multiple different effects on cellular form and function. Here, we review work implicating Redox intermediates in post-translationally altering actin and discuss what is known regarding how these alterations affect the properties of actin. We also focus on two of the best characterized enzymatic sources of these Redox intermediates – the NADPH oxidase NOX and the flavoprotein monooxygenase MICAL – and detail how they have both been identified as altering actin, but share little similarity and employ different means to regulate actin dynamics. Finally, we discuss the role of these enzymes and redox signaling in regulating the actin cytoskeleton in vivo and highlight their importance for neuronal form and function in health and disease. PMID:27309342

  5. Trauma ocupacional por corpo estranho corneano superficial Occupational trauma due to superficial corneal foreign body

    Directory of Open Access Journals (Sweden)

    Vanessa Miroski Gerente

    2008-04-01

    Full Text Available OBJETIVO: Avaliar a epidemiologia do trauma ocular por corpo estranho superficial de córnea. MÉTODOS: Os pacientes atendidos no Pronto-Socorro da Universidade Federal de São Paulo entre abril e junho de 2005 que apresentaram corpo estranho superficial de córnea foram entrevistados. Foram avaliados: sexo, idade, profissão, registro legal do emprego, uso, disponibilidade e tipo de equipamentos de proteção utilizados e a fiscalização do seu uso. O conhecimento das complicações deste tipo de acidente também foi avaliado. Os resultados foram analisados com teste do qui quadrado ou teste de Fisher. RESULTADOS: Foram entrevistados 123 pacientes. Apenas 3 eram do sexo feminino e a idade média foi de 36 anos. A maioria destes traumas ocorreu no ambiente de trabalho (86,2% e 58,4% não possuíam registro legal do emprego. As profissões mais freqüentemente envolvidas foram serralheiro, pedreiro e metalúrgico. Em 79,8% dos locais de trabalho havia equipamentos de proteção e 85,3% dos pacientes eram orientados a usá-los. Em 52,4% dos locais sua utilização era fiscalizada, mas apenas 34,2% usavam no momento do trauma. A utilização foi mais freqüente (p=0,008 e fiscalização mais presente (p=0,0415 entre pacientes com registro legal de emprego. Questionados sobre os riscos, 68,9% dos pacientes tinham consciência das complicações graves deste tipo de acidente. CONCLUSÃO: A maioria dos pacientes tem conhecimento sobre a gravidade do trauma ocular e este tipo de lesão ocorre mesmo em locais com equipamentos de proteção disponíveis, alguns deles até durante o seu uso. Os dados sugerem que enfoque maior da prevenção deve ser na fiscalização e utilização de equipamentos adequados.PURPOSE: To evaluate the epidemiology of superficial corneal foreign body. METHODS: Patients who were seen at the Emergency Service of the Federal University of São Paulo, from April/05 to June/05, were screened and those with superficial corneal

  6. Superficial herpes simplex virus wound infection following lung transplantation.

    Science.gov (United States)

    Karolak, Wojtek; Wojarski, Jacek; Zegleń, Sławomir; Ochman, Marek; Urlik, Maciej; Hudzik, Bartosz; Wozniak-Grygiel, Elzbieta; Maruszewski, Marcin

    2017-08-01

    Surgical site infections (SSIs) are infections of tissues, organs, or spaces exposed by surgeons during performance of an invasive procedure. SSIs are classified into superficial, which are limited to skin and subcutaneous tissues, and deep. The incidence of deep SSIs in lung transplant (LTx) patients is estimated at 5%. No reports have been published as to the incidence of superficial SSIs specifically in LTx patients. Common sense would dictate that the majority of superficial SSIs would be bacterial. Uncommonly, fungal SSIs may occur, and we believe that no reports exist as to the incidence of viral wound infections in LTx patients, or in any solid organ transplant patients. We report a de novo superficial wound infection with herpes simplex virus following lung transplantation, its possible source, treatment, and resolution. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  7. Imaging of the most frequent superficial soft-tissue sarcomas

    International Nuclear Information System (INIS)

    Morel, Melanie; Taieb, Sophie; Ceugnart, Luc; Penel, Nicolas; Mortier, Laurent; Vanseymortier, Luc; Robin, Y.M.; Gosset, Pierre; Cotten, Anne

    2011-01-01

    Superficial soft-tissue sarcomas are malignant mesenchymal tumors located within the cutaneous and/or subcutaneous layers. Most superficial soft-tissue sarcomas are low-grade tumors; yet, the risk of local recurrence is high, and initial wide surgery is the main prognostic factor. Some of these superficial sarcomas may grow, following an infiltrative pattern, and their real extent may be underestimated clinically. Imaging techniques are useful to determine precisely the real margins of the tumor, especially in cases of clinically doubtful or recurrent or large superficial lesions. Imaging tools enable one to determine the relationship with the superficial fascia separating the subcutaneous layer from the underlying muscle. In our institution ultrasonographic examination is followed by magnetic resonance (MR) imaging when the size of the lesion exceeds 3-5 cm. Imaging assessment is performed prior to biopsy, enabling optimal surgical management. Imaging features of the main superficial sarcomas are detailed in the following article, according to their major locations: those arising in the epidermis and/or dermis, which are most often diagnosed by dermatologists, and the subcutaneous sarcomas. (orig.)

  8. Imaging of the most frequent superficial soft-tissue sarcomas

    Energy Technology Data Exchange (ETDEWEB)

    Morel, Melanie; Taieb, Sophie; Ceugnart, Luc [Centre Oscar Lambret, Department of Radiology, Lille (France); Penel, Nicolas [Centre Oscar Lambret, Department of Oncology, Lille (France); Mortier, Laurent [Centre Hospitalier Universitaire de Lille, Department of Dermatology, Hopital Claude Huriez, Lille (France); Vanseymortier, Luc [Centre Oscar Lambret, Department of Surgery, Lille (France); Robin, Y.M. [Centre Oscar Lambret, Departement of Pathology, Lille (France); Gosset, Pierre [Groupement Hospitalier de l' Institut Catholique-Faculte Libre de Medecine de Lille, Department of Pathology, Hopital Saint-Philibert, Lomme (France); Cotten, Anne [Centre Hospitalier Universitaire de Lille, Department of Musculoskeletal Radiology, Centre Hopital Roger Salengro, Lille (France)

    2011-03-15

    Superficial soft-tissue sarcomas are malignant mesenchymal tumors located within the cutaneous and/or subcutaneous layers. Most superficial soft-tissue sarcomas are low-grade tumors; yet, the risk of local recurrence is high, and initial wide surgery is the main prognostic factor. Some of these superficial sarcomas may grow, following an infiltrative pattern, and their real extent may be underestimated clinically. Imaging techniques are useful to determine precisely the real margins of the tumor, especially in cases of clinically doubtful or recurrent or large superficial lesions. Imaging tools enable one to determine the relationship with the superficial fascia separating the subcutaneous layer from the underlying muscle. In our institution ultrasonographic examination is followed by magnetic resonance (MR) imaging when the size of the lesion exceeds 3-5 cm. Imaging assessment is performed prior to biopsy, enabling optimal surgical management. Imaging features of the main superficial sarcomas are detailed in the following article, according to their major locations: those arising in the epidermis and/or dermis, which are most often diagnosed by dermatologists, and the subcutaneous sarcomas. (orig.)

  9. Plasmin enzymatic activity in the presence of actin

    Directory of Open Access Journals (Sweden)

    Yusova E. I.

    2015-10-01

    Full Text Available Aim. To study the changes in the plasmin activity towards substrates with high and low molecular mass in the presence of actin. Methods. The proteins used for this investigation were obtained by affinity chromatography and gel-filtration. The plasmin enzymatic activity was determined by a turbidimetric assay and a chromogenic substrate-based assay. The enzyme linked immunosorbent assay and biotin-avidin-phosphatase system were used to study the interaction of plasminogen and its fragments with actin. Results. It was shown that G-actin causes 1.5-fold decrease in the rate of polymeric fibrin hydrolysis by plasmin and Glu-plasminogen activated by the tissue plasminogen activator. However, actin did not impede plasmin autolysis and had no influence on its amidase activity. We have studied an interaction of biotinylated Glu-plasminogen and its fragments (kringle 1-3, kringle 4 and mini-plasminogen with immobilized G-actin. Glu-plasminogen and kringle 4 had a high affinity towards actin (C50 is 113 and 117 nM correspondingly. Mini-plasminogen and kringe 4 did not bind to actin. A similar affinity of Glu-plasminogen and kringle 1-3 towards actin proves the involvement of the kringle 1-3 lysine-binding sites of the native plasminogen form in the actin interaction. Conclusions. Actin can modulate plasmin specificity towards high molecular mass substrates through its interaction with lysine-binding sites of the enzyme kringle domains. Actin inhibition of the fibrinolytic activity of plasmin is due to its competition with fibrin for thelysine binding sites of plasminogen/plasmin.

  10. The cell wall of Arabidopsis thaliana influences actin network dynamics.

    Science.gov (United States)

    Tolmie, Frances; Poulet, Axel; McKenna, Joseph; Sassmann, Stefan; Graumann, Katja; Deeks, Michael; Runions, John

    2017-07-20

    In plant cells, molecular connections link the cell wall-plasma membrane-actin cytoskeleton to form a continuum. It is hypothesized that the cell wall provides stable anchor points around which the actin cytoskeleton remodels. Here we use live cell imaging of fluorescently labelled marker proteins to quantify the organization and dynamics of the actin cytoskeleton and to determine the impact of disrupting connections within the continuum. Labelling of the actin cytoskeleton with green fluorescent protein (GFP)-fimbrin actin-binding domain 2 (FABD2) resulted in a network composed of fine filaments and thicker bundles that appeared as a highly dynamic remodelling meshwork. This differed substantially from the GFP-Lifeact-labelled network that appeared much more sparse with thick bundles that underwent 'simple movement', in which the bundles slightly change position, but in such a manner that the structure of the network was not substantially altered during the time of observation. Label-dependent differences in actin network morphology and remodelling necessitated development of two new image analysis techniques. The first of these, 'pairwise image subtraction', was applied to measurement of the more rapidly remodelling actin network labelled with GFP-FABD2, while the second, 'cumulative fluorescence intensity', was used to measure bulk remodelling of the actin cytoskeleton when labelled with GFP-Lifeact. In each case, these analysis techniques show that the actin cytoskeleton has a decreased rate of bulk remodelling when the cell wall-plasma membrane-actin continuum is disrupted either by plasmolysis or with isoxaben, a drug that specifically inhibits cellulose deposition. Changes in the rate of actin remodelling also affect its functionality, as observed by alteration in Golgi body motility. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  11. Morphodynamics of the Actin-Rich Cytoskeleton in Entamoeba histolytica

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    Maria Manich

    2018-05-01

    Full Text Available Entamoeba histolytica is the anaerobic protozoan parasite responsible for human amoebiasis, the third most deadly parasitic disease worldwide. This highly motile eukaryotic cell invades human tissues and constitutes an excellent experimental model of cell motility and cell shape deformation. The absence of extranuclear microtubules in Entamoeba histolytica means that the actin-rich cytoskeleton takes on a crucial role in not only amoebic motility but also other processes sustaining pathogenesis, such as the phagocytosis of human cells and the parasite's resistance of host immune responses. Actin is highly conserved among eukaryotes, although diverse isoforms exist in almost all organisms studied to date. However, E. histolytica has a single actin protein, the structure of which differs significantly from those of its human homologs. Here, we studied the expression, structure and dynamics of actin in E. histolytica. We used molecular and cellular approaches to evaluate actin gene expression during intestinal invasion by E. histolytica trophozoites. Based on a three-dimensional structural bioinformatics analysis, we characterized protein domains differences between amoebic actin and human actin. Fine-tuned molecular dynamics simulations enabled us to examine protein motion and refine the three-dimensional structures of both actins, including elements potentially accounting for differences changes in the affinity properties of amoebic actin and deoxyribonuclease I. The dynamic, multifunctional nature of the amoebic cytoskeleton prompted us to examine the pleiotropic forms of actin structures within live E. histolytica cells; we observed the cortical cytoskeleton, stress fibers, “dot-like” structures, adhesion plates, and macropinosomes. In line with these data, a proteomics study of actin-binding proteins highlighted the Arp2/3 protein complex as a crucial element for the development of macropinosomes and adhesion plaques.

  12. Actin and Endocytosis in Budding Yeast

    Science.gov (United States)

    Goode, Bruce L.; Eskin, Julian A.; Wendland, Beverly

    2015-01-01

    Endocytosis, the process whereby the plasma membrane invaginates to form vesicles, is essential for bringing many substances into the cell and for membrane turnover. The mechanism driving clathrin-mediated endocytosis (CME) involves > 50 different protein components assembling at a single location on the plasma membrane in a temporally ordered and hierarchal pathway. These proteins perform precisely choreographed steps that promote receptor recognition and clustering, membrane remodeling, and force-generating actin-filament assembly and turnover to drive membrane invagination and vesicle scission. Many critical aspects of the CME mechanism are conserved from yeast to mammals and were first elucidated in yeast, demonstrating that it is a powerful system for studying endocytosis. In this review, we describe our current mechanistic understanding of each step in the process of yeast CME, and the essential roles played by actin polymerization at these sites, while providing a historical perspective of how the landscape has changed since the preceding version of the YeastBook was published 17 years ago (1997). Finally, we discuss the key unresolved issues and where future studies might be headed. PMID:25657349

  13. Drenaje suplementario del sistema venoso superficial en colgajos pediculados Supplementary drainage of superficial venous system in pedicled flaps

    Directory of Open Access Journals (Sweden)

    A. Fernández García

    2012-06-01

    Full Text Available Los colgajos pediculados pueden sufrir edema y congestión debido a que su drenaje a trevés del sistema venoso superfical es imposible y el flujo hacia el sistema profundo a nivel del pedículo es precario. Esta situación suele evolucionar hacia la necrosis parcial o la pérdida total del colgajo en pocas horas. La apertura del sistema venoso superficial permite el drenaje adecuado de los tejidos transferidos y evita estas complicaciones. Este artículo analiza el papel de las anastomosis microquirúrgicas en el sistema venoso superficial de los colgajos pediculados. Esta técnica permite comunicar el sistema venoso superfical de los tejidos transferidos con el sistema venoso superficial del territorio que rodea al defecto. Presentamos los resultados obtenidos con esta técnica en colgajos de perforante en hélice, colgajos miocutáneos, colgajos neurocutáneos y colgajos fasciograsos volteados.The pedicled flaps can suffer edema and congestion due to the impossibility of drainage toward the superficial venous system and the precarious flow via the deep system at the level of the pedicle. The evolution of this situation is usually partial necrosis or total loss of the flap in a few hours. The opening of the venous superficial system achieves an appropriate drainage of the transfered tissues and avoids these complications. In this article we analyze the role of the microsurgical anastomosis at the level of the superficial venous system of pedicled flaps. This technique allows to communicate the superficial venous system of the transfered tissues with the superficial venous system of the territory around the defect. We report the results with this method in propeller perforator flaps, miocutaneous flaps, neurocutaneous flaps and adipofascial turn over flaps.

  14. A Legionella Effector Disrupts Host Cytoskeletal Structure by Cleaving Actin.

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    Yao Liu

    2017-01-01

    Full Text Available Legionella pneumophila, the etiological agent of Legionnaires' disease, replicates intracellularly in protozoan and human hosts. Successful colonization and replication of this pathogen in host cells requires the Dot/Icm type IVB secretion system, which translocates approximately 300 effector proteins into the host cell to modulate various cellular processes. In this study, we identified RavK as a Dot/Icm substrate that targets the host cytoskeleton and reduces actin filament abundance in mammalian cells upon ectopic expression. RavK harbors an H95EXXH99 motif associated with diverse metalloproteases, which is essential for the inhibition of yeast growth and for the induction of cell rounding in HEK293T cells. We demonstrate that the actin protein itself is the cellular target of RavK and that this effector cleaves actin at a site between residues Thr351 and Phe352. Importantly, RavK-mediated actin cleavage also occurs during L. pneumophila infection. Cleavage by RavK abolishes the ability of actin to form polymers. Furthermore, an F352A mutation renders actin resistant to RavK-mediated cleavage; expression of the mutant in mammalian cells suppresses the cell rounding phenotype caused by RavK, further establishing that actin is the physiological substrate of RavK. Thus, L. pneumophila exploits components of the host cytoskeleton by multiple effectors with distinct mechanisms, highlighting the importance of modulating cellular processes governed by the actin cytoskeleton in the intracellular life cycle of this pathogen.

  15. Apatite-mediated actin dynamics in resorbing osteoclasts.

    Science.gov (United States)

    Saltel, Frédéric; Destaing, Olivier; Bard, Frédéric; Eichert, Diane; Jurdic, Pierre

    2004-12-01

    The actin cytoskeleton is essential for osteoclasts main function, bone resorption. Two different organizations of actin have been described in osteoclasts, the podosomes belt corresponding to numerous F-actin columns arranged at the cell periphery, and the sealing zone defined as a unique large band of actin. To compare the role of these two different actin organizations, we imaged osteoclasts on various substrata: glass, dentin, and apatite. Using primary osteoclasts expressing GFP-actin, we found that podosome belts and sealing zones, both very dynamic actin structures, were present in mature osteoclasts; podosome belts were observed only in spread osteoclasts adhering onto glass, whereas sealing zone were seen in apico-basal polarized osteoclasts adherent on mineralized matrix. Dynamic observations of several resorption cycles of osteoclasts seeded on apatite revealed that 1) podosomes do not fuse together to form the sealing zone; 2) osteoclasts alternate successive stationary polarized resorption phases with a sealing zone and migration, nonresorption phases without any specific actin structure; and 3) apatite itself promotes sealing zone formation though c-src and Rho signaling. Finally, our work suggests that apatite-mediated sealing zone formation is dependent on both c-src and Rho whereas apico-basal polarization requires only Rho.

  16. The evolution of compositionally and functionally distinct actin filaments.

    Science.gov (United States)

    Gunning, Peter W; Ghoshdastider, Umesh; Whitaker, Shane; Popp, David; Robinson, Robert C

    2015-06-01

    The actin filament is astonishingly well conserved across a diverse set of eukaryotic species. It has essentially remained unchanged in the billion years that separate yeast, Arabidopsis and man. In contrast, bacterial actin-like proteins have diverged to the extreme, and many of them are not readily identified from sequence-based homology searches. Here, we present phylogenetic analyses that point to an evolutionary drive to diversify actin filament composition across kingdoms. Bacteria use a one-filament-one-function system to create distinct filament systems within a single cell. In contrast, eukaryotic actin is a universal force provider in a wide range of processes. In plants, there has been an expansion of the number of closely related actin genes, whereas in fungi and metazoa diversification in tropomyosins has increased the compositional variety in actin filament systems. Both mechanisms dictate the subset of actin-binding proteins that interact with each filament type, leading to specialization in function. In this Hypothesis, we thus propose that different mechanisms were selected in bacteria, plants and metazoa, which achieved actin filament compositional variation leading to the expansion of their functional diversity. © 2015. Published by The Company of Biologists Ltd.

  17. Actin is an essential component of plant gravitropic signaling pathways

    Science.gov (United States)

    Braun, Markus; Hauslage, Jens; Limbach, Christoph

    2003-08-01

    A role of the actin cytoskeleton in the different phases of gravitropism in higher plant organs seems obvious, but experimental evidence is still inconclusive and contradictory. In gravitropically tip-growing rhizoids and protonemata, however, it is well documented that actin is an essential component of the tip-growth machinery and is involved either in the cellular mechanisms that lead to gravity sensing and in the processes of the graviresponses that result in the reorientation of the growth direction. All these processes depend on a complexly organized and highly dynamic organization of actin filaments whose diverse functions are coordinated by numerous associated proteins. Actin filaments and myosins mediate the transport of secretory vehicles to the growing tip and precisely control the delivery of cell wall material. In addition, both cell types use a very efficient actomyosin-based system to control and correct the position of their statoliths and to direct sedimenting statoliths to confined graviperception sites at the plasma membrane. The studies presented in this paper provide evidence for the essential role of actin in plant gravity sensing and the gravitropic responses. A unique actin-organizing center exists in the tip of characean rhizoids and protonemata which is associated with and dynamically regulated by a specific set of actin-dynamizing proteins. It is concluded that this highly dynamic apical actin array is an essential prerequisite for gravity sensing and gravity-oriented tip growth.

  18. Stochastic Severing of Actin Filaments by Actin Depolymerizing Factor/Cofilin Controls the Emergence of a Steady Dynamical Regime

    Science.gov (United States)

    Roland, Jeremy; Berro, Julien; Michelot, Alphée; Blanchoin, Laurent; Martiel, Jean-Louis

    2008-01-01

    Actin dynamics (i.e., polymerization/depolymerization) powers a large number of cellular processes. However, a great deal remains to be learned to explain the rapid actin filament turnover observed in vivo. Here, we developed a minimal kinetic model that describes key details of actin filament dynamics in the presence of actin depolymerizing factor (ADF)/cofilin. We limited the molecular mechanism to 1), the spontaneous growth of filaments by polymerization of actin monomers, 2), the ageing of actin subunits in filaments, 3), the cooperative binding of ADF/cofilin to actin filament subunits, and 4), filament severing by ADF/cofilin. First, from numerical simulations and mathematical analysis, we found that the average filament length, 〈L〉, is controlled by the concentration of actin monomers (power law: 5/6) and ADF/cofilin (power law: −2/3). We also showed that the average subunit residence time inside the filament, 〈T〉, depends on the actin monomer (power law: −1/6) and ADF/cofilin (power law: −2/3) concentrations. In addition, filament length fluctuations are ∼20% of the average filament length. Moreover, ADF/cofilin fragmentation while modulating filament length keeps filaments in a high molar ratio of ATP- or ADP-Pi versus ADP-bound subunits. This latter property has a protective effect against a too high severing activity of ADF/cofilin. We propose that the activity of ADF/cofilin in vivo is under the control of an affinity gradient that builds up dynamically along growing actin filaments. Our analysis shows that ADF/cofilin regulation maintains actin filaments in a highly dynamical state compatible with the cytoskeleton dynamics observed in vivo. PMID:18065447

  19. Actin dynamics, architecture, and mechanics in cell motility.

    Science.gov (United States)

    Blanchoin, Laurent; Boujemaa-Paterski, Rajaa; Sykes, Cécile; Plastino, Julie

    2014-01-01

    Tight coupling between biochemical and mechanical properties of the actin cytoskeleton drives a large range of cellular processes including polarity establishment, morphogenesis, and motility. This is possible because actin filaments are semi-flexible polymers that, in conjunction with the molecular motor myosin, can act as biological active springs or "dashpots" (in laymen's terms, shock absorbers or fluidizers) able to exert or resist against force in a cellular environment. To modulate their mechanical properties, actin filaments can organize into a variety of architectures generating a diversity of cellular organizations including branched or crosslinked networks in the lamellipodium, parallel bundles in filopodia, and antiparallel structures in contractile fibers. In this review we describe the feedback loop between biochemical and mechanical properties of actin organization at the molecular level in vitro, then we integrate this knowledge into our current understanding of cellular actin organization and its physiological roles.

  20. Toward the Structure of Dynamic Membrane-Anchored Actin Networks

    Science.gov (United States)

    Weber, Igor

    2007-01-01

    In the cortex of a motile cell, membrane-anchored actin filaments assemble into structures of varying shape and function. Filopodia are distinguished by a core of bundled actin filaments within finger-like extensions of the membrane. In a recent paper by Medalia et al1 cryo-electron tomography has been used to reconstruct, from filopodia of Dictyostelium cells, the 3-dimensional organization of actin filaments in connection with the plasma membrane. A special arrangement of short filaments converging toward the filopod's tip has been called a “terminal cone”. In this region force is applied for protrusion of the membrane. Here we discuss actin organization in the filopodia of Dictyostelium in the light of current views on forces that are generated by polymerizing actin filaments, and on the resistance of membranes against deformation that counteracts these forces. PMID:19262130

  1. Initiation of DNA replication requires actin dynamics and formin activity.

    Science.gov (United States)

    Parisis, Nikolaos; Krasinska, Liliana; Harker, Bethany; Urbach, Serge; Rossignol, Michel; Camasses, Alain; Dewar, James; Morin, Nathalie; Fisher, Daniel

    2017-11-02

    Nuclear actin regulates transcriptional programmes in a manner dependent on its levels and polymerisation state. This dynamics is determined by the balance of nucleocytoplasmic shuttling, formin- and redox-dependent filament polymerisation. Here, using Xenopus egg extracts and human somatic cells, we show that actin dynamics and formins are essential for DNA replication. In proliferating cells, formin inhibition abolishes nuclear transport and initiation of DNA replication, as well as general transcription. In replicating nuclei from transcriptionally silent Xenopus egg extracts, we identified numerous actin regulators, and disruption of actin dynamics abrogates nuclear transport, preventing NLS (nuclear localisation signal)-cargo release from RanGTP-importin complexes. Nuclear formin activity is further required to promote loading of cyclin-dependent kinase (CDK) and proliferating cell nuclear antigen (PCNA) onto chromatin, as well as initiation and elongation of DNA replication. Therefore, actin dynamics and formins control DNA replication by multiple direct and indirect mechanisms. © 2017 The Authors.

  2. A nucleator arms race: cellular control of actin assembly.

    Science.gov (United States)

    Campellone, Kenneth G; Welch, Matthew D

    2010-04-01

    For over a decade, the actin-related protein 2/3 (ARP2/3) complex, a handful of nucleation-promoting factors and formins were the only molecules known to directly nucleate actin filament formation de novo. However, the past several years have seen a surge in the discovery of mammalian proteins with roles in actin nucleation and dynamics. Newly recognized nucleation-promoting factors, such as WASP and SCAR homologue (WASH), WASP homologue associated with actin, membranes and microtubules (WHAMM), and junction-mediating regulatory protein (JMY), stimulate ARP2/3 activity at distinct cellular locations. Formin nucleators with additional biochemical and cellular activities have also been uncovered. Finally, the Spire, cordon-bleu and leiomodin nucleators have revealed new ways of overcoming the kinetic barriers to actin polymerization.

  3. Clinical Response to Ingenol Mebutate in Patients With Actinic Keratoses.

    Science.gov (United States)

    Batalla, A; Flórez, Á; Feal, C; Peón, G; Abalde, M T; Salgado-Boquete, L; de la Torre, C

    2015-12-01

    Cryotherapy is the most common treatment for actinic keratosis, but its effect is limited to individual lesions. Several topical drugs, however, are available that, in addition to treating individual actinic keratoses, target field cancerization and thereby act on subclinical lesions. Examples are 5-fluorouracil, imiquimod, diclofenac, and ingenol mebutate. We report on 17 patients with actinic keratoses treated with ingenol mebutate and describe our findings on treatment effectiveness, adherence, and tolerance. Complete and partial response rates were 35% and 53%, respectively. Ninety-four percent of patients fully adhered to treatment and 18% developed severe local reactions. Ingenol mebutate is an effective treatment for actinic keratosis. Although it has a similar rate of local reactions to other treatments available for actinic keratosis, its short treatment regimen favors better adherence. Copyright © 2014 Elsevier España, S.L.U. y AEDV. All rights reserved.

  4. Extracellular Actin Is a Receptor for Mycoplasma hyopneumoniae

    Directory of Open Access Journals (Sweden)

    Benjamin B. A. Raymond

    2018-02-01

    Full Text Available Mycoplasma hyopneumoniae, an agriculturally important porcine pathogen, disrupts the mucociliary escalator causing ciliostasis, loss of cilial function, and epithelial cell death within the porcine lung. Losses to swine production due to growth rate retardation and reduced feed conversion efficiency are severe, and antibiotics are used heavily to control mycoplasmal pneumonia. Notably, little is known about the repertoire of host receptors that M. hyopneumoniae targets to facilitate colonization. Here we show, for the first time, that actin exists extracellularly on porcine epithelial monolayers (PK-15 using surface biotinylation and 3D-Structured Illumination Microscopy (3D-SIM, and that M. hyopneumoniae binds to the extracellular β-actin exposed on the surface of these cells. Consistent with this hypothesis we show: (i monoclonal antibodies that target β-actin significantly block the ability of M. hyopneumoniae to adhere and colonize PK-15 cells; (ii microtiter plate binding assays show that M. hyopneumoniae cells bind to monomeric G-actin in a dose dependent manner; (iii more than 100 M. hyopneumoniae proteins were recovered from affinity-chromatography experiments using immobilized actin as bait; and (iv biotinylated monomeric actin binds directly to M. hyopneumoniae proteins in ligand blotting studies. Specifically, we show that the P97 cilium adhesin possesses at least two distinct actin-binding regions, and binds monomeric actin with nanomolar affinity. Taken together, these observations suggest that actin may be an important receptor for M. hyopneumoniae within the swine lung and will aid in the future development of intervention strategies against this devastating pathogen. Furthermore, our observations have wider implications for extracellular actin as an important bacterial receptor.

  5. Extracellular Actin Is a Receptor for Mycoplasma hyopneumoniae.

    Science.gov (United States)

    Raymond, Benjamin B A; Madhkoor, Ranya; Schleicher, Ina; Uphoff, Cord C; Turnbull, Lynne; Whitchurch, Cynthia B; Rohde, Manfred; Padula, Matthew P; Djordjevic, Steven P

    2018-01-01

    Mycoplasma hyopneumoniae , an agriculturally important porcine pathogen, disrupts the mucociliary escalator causing ciliostasis, loss of cilial function, and epithelial cell death within the porcine lung. Losses to swine production due to growth rate retardation and reduced feed conversion efficiency are severe, and antibiotics are used heavily to control mycoplasmal pneumonia. Notably, little is known about the repertoire of host receptors that M. hyopneumoniae targets to facilitate colonization. Here we show, for the first time, that actin exists extracellularly on porcine epithelial monolayers (PK-15) using surface biotinylation and 3D-Structured Illumination Microscopy (3D-SIM), and that M. hyopneumoniae binds to the extracellular β-actin exposed on the surface of these cells. Consistent with this hypothesis we show: (i) monoclonal antibodies that target β-actin significantly block the ability of M. hyopneumoniae to adhere and colonize PK-15 cells; (ii) microtiter plate binding assays show that M. hyopneumoniae cells bind to monomeric G-actin in a dose dependent manner; (iii) more than 100 M. hyopneumoniae proteins were recovered from affinity-chromatography experiments using immobilized actin as bait; and (iv) biotinylated monomeric actin binds directly to M. hyopneumoniae proteins in ligand blotting studies. Specifically, we show that the P97 cilium adhesin possesses at least two distinct actin-binding regions, and binds monomeric actin with nanomolar affinity. Taken together, these observations suggest that actin may be an important receptor for M. hyopneumoniae within the swine lung and will aid in the future development of intervention strategies against this devastating pathogen. Furthermore, our observations have wider implications for extracellular actin as an important bacterial receptor.

  6. IFT88 influences chondrocyte actin organization and biomechanics.

    Science.gov (United States)

    Wang, Z; Wann, A K T; Thompson, C L; Hassen, A; Wang, W; Knight, M M

    2016-03-01

    Primary cilia are microtubule based organelles which control a variety of signalling pathways important in cartilage development, health and disease. This study examines the role of the intraflagellar transport (IFT) protein, IFT88, in regulating fundamental actin organisation and mechanics in articular chondrocytes. The study used an established chondrocyte cell line with and without hypomorphic mutation of IFT88 (IFT88(orpk)). Confocal microscopy was used to quantify F-actin and myosin IIB organisation. Viscoelastic cell and actin cortex mechanics were determined using micropipette aspiration with actin dynamics visualised in live cells transfected with LifeACT-GFP. IFT88(orpk) cells exhibited a significant increase in acto-myosin stress fibre organisation relative to wild-type (WT) cells in monolayer and an altered response to cytochalasin D. Rounded IFT88(orpk) cells cultured in suspension exhibited reduced cortical actin expression with reduced cellular equilibrium modulus. Micropipette aspiration resulted in reduced membrane bleb formation in IFT88(orpk) cells. Following membrane blebbing, IFT88(orpk) cells exhibited slower reformation of the actin cortex. IFT88(orpk) cells showed increased actin deformability and reduced cortical tension confirming that IFT regulates actin cortex mechanics. The reduced cortical tension is also consistent with the reduced bleb formation. This study demonstrates for the first time that the ciliary protein IFT88 regulates fundamental actin organisation and the stiffness of the actin cortex leading to alterations in cell deformation, mechanical properties and blebbing in an IFT88 chondrocyte cell line. This adds to the growing understanding of the role of primary cilia and IFT in regulating cartilage biology. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  7. [Actinic keratosis: New concept and therapeutic update].

    Science.gov (United States)

    Carmena-Ramón, Rafael; Mateu-Puchades, Almudena; Santos-Alarcón, Sergio; Lucas-Truyols, Sofía

    2017-10-01

    Actinic keratosis (AK) is a common reason for consultation in both Primary Care and Specialised Care. It is the third or fourth most common reason for consultation in dermatology, accounting for up to 5-6% of patients attended. It has also been observed that its prevalence has been increasing in the last 10years, compared to other dermatoses. This is also expected to continue to increase due to longer life expectancy, and by the changes in sun exposure habits since the middle of the last century. The aim of this article is to update the concepts of AK, cancerisation field and to present the currently available therapeutic tools. Copyright © 2017. Publicado por Elsevier España, S.L.U.

  8. Superficial dose evaluation of four dose calculation algorithms

    Science.gov (United States)

    Cao, Ying; Yang, Xiaoyu; Yang, Zhen; Qiu, Xiaoping; Lv, Zhiping; Lei, Mingjun; Liu, Gui; Zhang, Zijian; Hu, Yongmei

    2017-08-01

    Accurate superficial dose calculation is of major importance because of the skin toxicity in radiotherapy, especially within the initial 2 mm depth being considered more clinically relevant. The aim of this study is to evaluate superficial dose calculation accuracy of four commonly used algorithms in commercially available treatment planning systems (TPS) by Monte Carlo (MC) simulation and film measurements. The superficial dose in a simple geometrical phantom with size of 30 cm×30 cm×30 cm was calculated by PBC (Pencil Beam Convolution), AAA (Analytical Anisotropic Algorithm), AXB (Acuros XB) in Eclipse system and CCC (Collapsed Cone Convolution) in Raystation system under the conditions of source to surface distance (SSD) of 100 cm and field size (FS) of 10×10 cm2. EGSnrc (BEAMnrc/DOSXYZnrc) program was performed to simulate the central axis dose distribution of Varian Trilogy accelerator, combined with measurements of superficial dose distribution by an extrapolation method of multilayer radiochromic films, to estimate the dose calculation accuracy of four algorithms in the superficial region which was recommended in detail by the ICRU (International Commission on Radiation Units and Measurement) and the ICRP (International Commission on Radiological Protection). In superficial region, good agreement was achieved between MC simulation and film extrapolation method, with the mean differences less than 1%, 2% and 5% for 0°, 30° and 60°, respectively. The relative skin dose errors were 0.84%, 1.88% and 3.90%; the mean dose discrepancies (0°, 30° and 60°) between each of four algorithms and MC simulation were (2.41±1.55%, 3.11±2.40%, and 1.53±1.05%), (3.09±3.00%, 3.10±3.01%, and 3.77±3.59%), (3.16±1.50%, 8.70±2.84%, and 18.20±4.10%) and (14.45±4.66%, 10.74±4.54%, and 3.34±3.26%) for AXB, CCC, AAA and PBC respectively. Monte Carlo simulation verified the feasibility of the superficial dose measurements by multilayer Gafchromic films. And the rank

  9. Titin-Actin Interaction: PEVK-Actin-Based Viscosity in a Large Animal

    Directory of Open Access Journals (Sweden)

    Charles S. Chung

    2011-01-01

    Full Text Available Titin exhibits an interaction between its PEVK segment and the actin filament resulting in viscosity, a speed dependent resistive force, which significantly influences diastolic filling in mice. While diastolic disease is clinically pervasive, humans express a more compliant titin (N2BA:N2B ratio ~0.5–1.0 than mice (N2BA:N2B ratio ~0.2. To examine PEVK-actin based viscosity in compliant titin-tissues, we used pig cardiac tissue that expresses titin isoforms similar to that in humans. Stretch-hold experiments were performed at speeds from 0.1 to 10 lengths/s from slack sarcomere lengths (SL to SL of 2.15 μm. Viscosity was calculated from the slope of stress-relaxation vs stretch speed. Recombinant PEVK was added to compete off native interactions and this found to reduce the slope by 35%, suggesting that PEVK-actin interactions are a strong contributor of viscosity. Frequency sweeps were performed at frequencies of 0.1–400 Hz and recombinant protein reduced viscous moduli by 40% at 2.15 μm and by 50% at 2.25 μm, suggesting a SL-dependent nature of viscosity that might prevent SL ``overshoot’’ at long diastolic SLs. This study is the first to show that viscosity is present at physiologic speeds in the pig and supports the physiologic relevance of PEVK-actin interactions in humans in both health and disease.

  10. Ultrastructural localization of actin and actin-binding proteins in the nucleus

    Czech Academy of Sciences Publication Activity Database

    Dingová, Hana; Fukalová, Jana; Maninová, Miloslava; Philimonenko, Vlada; Hozák, Pavel

    2009-01-01

    Roč. 131, č. 3 (2009), s. 425-434 ISSN 0948-6143 R&D Projects: GA MŠk LC545 Grant - others:MŠk(CZ) LC06063 Program:LC Institutional research plan: CEZ:AV0Z50520514 Keywords : nuclear actin * ultrastructure * actin–binding proteins Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.021, year: 2009

  11. Long-term evolution of superficial optic disc drusen

    DEFF Research Database (Denmark)

    Larsen, Lasse Malmqvist; Lund-Andersen, Henrik; Hamann, Steffen

    2017-01-01

    with hereditary ODD were included. Results: Mean age at initial and follow-up examination was, respectively, 16.8 and 73.3 years. The median follow-up time was 56 years. A minimal or non-existing change in superficial ODD anatomy (size and number) was seen in 10 of 12 eyes. There was a tendency towards more......Purpose: Optic disc drusen (ODD) is hyaline deposits in the optic nerve head seen in 1-2% of the population. Long-term evolution of ODD anatomy and visual field defects in ODD patients is a key factor for learning more about pathophysiology and prognosis of the condition. With a median follow......-up period of 56 years, this is the first study that evaluates superficial optic disc anatomy and visual fields in patients with ODD over a life span. Methods: Observational case series investigating progression of superficial optic disc anatomy and visual fields in ODD patients. Eight patients...

  12. Pigmented basal cell carcinoma mimicking a superficial spreading melanoma

    Directory of Open Access Journals (Sweden)

    Paula Hasbún Acuña

    2016-12-01

    Full Text Available Resumen El carcinoma basocelular es el cáncer de piel más frecuente, especialmente en personas de edad avanzada. El carcinoma basocelular pigmentado es una variante poco común que se ha descrito en la literatura como una lesión nodular hiperpigmentada. En raras ocasiones puede presentarse en forma de una extensa placa pigmentada, la cual puede ser clínicamente indistinguible del melanoma maligno de extensión superficial y de la enfermedad de Bowen. La dermatoscopía tiene una alta sensibilidad en el diagnóstico del carcinoma basocelular, cuando se utilizan los criterios de Menzies, aunque el diagnóstico final es histopatológico. El objetivo del presente trabajo es reportar y analizar el caso de una paciente con un extenso carcinoma basocelular superficial pigmentado, que simula un melanoma maligno de extensión superficial.

  13. Superficial siderosis of the central nervous system - A case report

    International Nuclear Information System (INIS)

    Mannalini, S.

    1997-01-01

    There is little information on superficial siderosis of the central nervous system (CNS) in the literature, mainly due to the rarity of the disease, the difficulties in diagnosis (autopsy pre magnetic resonance imaging (MRI)) and the long latency of the symptoms. With the advent of MRI, for the first time we are able to make a positive in vivo diagnosis. But this comes at a time of less disease incidence, and little clinical awareness. MRI is able to make the diagnosis because of the strong paramagnetic effect of haemosiderin, the blood by-product that is deposited on the brain surface in superficial siderosis of the CNS. The ability of the brain to biosynthesize ferritin in response to prolonged contact with haemosiderin is thought to be the most important factor in the pathogenesis of superficial siderosis. (author)

  14. Crosstalk between Rac1-mediated actin regulation and ROS production.

    Science.gov (United States)

    Acevedo, Alejandro; González-Billault, Christian

    2018-02-20

    The small RhoGTPase Rac1 is implicated in a variety of events related to actin cytoskeleton rearrangement. Remarkably, another event that is completely different from those related to actin regulation has the same relevance; the Rac1-mediated production of reactive oxygen species (ROS) through NADPH oxidases (NOX). Each outcome involves different Rac1 downstream effectors; on one hand, events related to the actin cytoskeleton require Rac1 to bind to WAVEs proteins and PAKs that ultimately promote actin branching and turnover, on the other, NOX-derived ROS production demands active Rac1 to be bound to a cytosolic activator of NOX. How Rac1-mediated signaling ends up promoting actin-related events, NOX-derived ROS, or both is poorly understood. Rac1 regulators, including scaffold proteins, are known to exert tight control over its functions. Hence, evidence of Rac1 regulatory events leading to both actin remodeling and NOX-mediated ROS generation are discussed. Moreover, cellular functions linked to physiological and pathological conditions that exhibit crosstalk between Rac1 outcomes are analyzed, while plausible roles in neuronal functions (and dysfunctions) are highlighted. Together, discussed evidence shed light on cellular mechanisms which requires Rac1 to direct either actin- and/or ROS-related events, helping to understand crucial roles of Rac1 dual functionality. Copyright © 2018 Elsevier Inc. All rights reserved.

  15. Biphasic interactions between a cationic dendrimer and actin.

    Science.gov (United States)

    Ruenraroengsak, Pakatip; Florence, Alexander T

    2010-12-01

    Gene delivery systems face the problem not only of the route toward the cell and tissues in question, but also of the molecularly crowded environment of both the cytoplasm and the nucleus itself. One of the physical barriers in the cytoplasm for diffusing nanoparticles is an actin network. Here, we describe the finding that a self-fluorescent sixth generation cationic dendrimer (6 nm in diameter) interacts reversibly and possibly electrostatically with actin filaments in vitro. Not only does this interaction slow the diffusion of the dendrimer but it also affects actin polymerization in a biphasic manner. At low concentrations the dendrimer behaves like a G-binding actin protein, retarding actin polymerization, whereas at high concentrations the dendrimer acts as a nucleating protein accelerating the polymerization. Thus in vivo the diffusion of a dendrimer carrier such as this has both physical and chemical elements: by decreasing polymerization it might accelerate its own transport, and by enhancing actin polymerization retard it. This finding suggests that such a dendrimer may have a role as an anticancer agent through its inhibitory effect on actin polymerization.

  16. Developmental expression of the alpha-skeletal actin gene

    Directory of Open Access Journals (Sweden)

    Vonk Freek J

    2008-06-01

    Full Text Available Abstract Background Actin is a cytoskeletal protein which exerts a broad range of functions in almost all eukaryotic cells. In higher vertebrates, six primary actin isoforms can be distinguished: alpha-skeletal, alpha-cardiac, alpha-smooth muscle, gamma-smooth muscle, beta-cytoplasmic and gamma-cytoplasmic isoactin. Expression of these actin isoforms during vertebrate development is highly regulated in a temporal and tissue-specific manner, but the mechanisms and the specific differences are currently not well understood. All members of the actin multigene family are highly conserved, suggesting that there is a high selective pressure on these proteins. Results We present here a model for the evolution of the genomic organization of alpha-skeletal actin and by molecular modeling, illustrate the structural differences of actin proteins of different phyla. We further describe and compare alpha-skeletal actin expression in two developmental stages of five vertebrate species (mouse, chicken, snake, salamander and fish. Our findings confirm that alpha-skeletal actin is expressed in skeletal muscle and in the heart of all five species. In addition, we identify many novel non-muscular expression domains including several in the central nervous system. Conclusion Our results show that the high sequence homology of alpha-skeletal actins is reflected by similarities of their 3 dimensional protein structures, as well as by conserved gene expression patterns during vertebrate development. Nonetheless, we find here important differences in 3D structures, in gene architectures and identify novel expression domains for this structural and functional important gene.

  17. Characterizing interaction forces between actin and proteins of the tropomodulin family reveals the presence of the N-terminal actin-binding site in leiomodin.

    Science.gov (United States)

    Arslan, Baran; Colpan, Mert; Gray, Kevin T; Abu-Lail, Nehal I; Kostyukova, Alla S

    2018-01-15

    Tropomodulin family of proteins includes several isoforms of tropomodulins (Tmod) and leiomodins (Lmod). These proteins can sequester actin monomers or nucleate actin polymerization. Although it is known that their actin-binding properties are isoform-dependent, knowledge on how they vary in strengths of interactions with G-actin is missing. While it is confirmed in many studies that Tmods have two actin-binding sites, information on number and location of actin-binding sites in Lmod2 is controversial. We used atomic force microscopy to study interactions between G-actin and proteins of the tropomodulin family. Unbinding forces between G-actin and Tmod1, Tmod2, Tmod3, or Lmod2 were quantified. Our results indicated that Tmod1 and Tmod3 had unimodal force distributions, Tmod2 had a bimodal distribution and Lmod2 had a trimodal distribution. The number of force distributions correlates with the proteins' abilities to sequester actin or to nucleate actin polymerization. We assigned specific unbinding forces to the individual actin-binding sites of Tmod2 and Lmod2 using mutations that destroy actin-binding sites of Tmod2 and truncated Lmod2. Our results confirm the existence of the N-terminal actin-binding site in Lmod2. Altogether, our data demonstrate how the differences between the number and the strength of actin-binding sites of Tmod or Lmod translate to their functional abilities. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. Experiments on the superficial irradiation of spherical vegetables and fruits

    International Nuclear Information System (INIS)

    Kalman, B.; Kiraly, Z.

    1974-01-01

    A revolving facility was made for the homogeneous superficial irradiation of spherical vegetables and fruits (apricot, peach, tomato, apple, etc.) with fast electrons. After the building of the technological apparatus described in detail, dosimetrical measurements were carried out by a Van de Graff generator of 2 MV and it was proved, that the superficial irradiation had a smaller effect on the quality of the fresh fruits, than of the stored ones. The developed apparatus can be altered according to the ripe-rate of the products. (K.A.)

  19. Aneurysm of the superficial femoral artery in an infant

    Energy Technology Data Exchange (ETDEWEB)

    Watanabe, M.; Komuro, H.; Matoba, K.; Kaneko, M. [Dept. of Paediatric Surgery, Inst. of Clinical Medicine, Univ. of Tsukuba, Ibaraki (Japan); Niitsu, M.; Itai, Y. [Dept. of Radiology, Inst. of Clinical Medicine, Univ. of Tsukuba, Ibaraki (Japan)

    2003-04-01

    An isolated arterial aneurysm in childhood is extremely rare. We report a 1-year-old girl with an aneurysm of the right superficial femoral artery, presenting as an asymptomatic mass of the thigh. The aneurysm involved the whole superficial femoral artery (9 cm in length), and surgical treatment would have required replacement of the affected artery. Conservative treatment was chosen, influenced by the patient's rapid growth at that time. Non-invasive, 3-D contrast-enhanced magnetic resonance angiography (MRA) was useful as an alternative to conventional angiography for detailed evaluation of the femoral arteries, including the aneurysm. (orig.)

  20. Actin depolymerization enhances adipogenic differentiation in human stromal stem cells

    DEFF Research Database (Denmark)

    Chen, Li; Hu, Huimin; Qiu, Weimin

    2018-01-01

    Human stromal stem cells (hMSCs) differentiate into adipocytes that play a role in skeletal tissue homeostasis and whole body energy metabolism. During adipocyte differentiation, hMSCs exhibit significant changes in cell morphology suggesting changes in cytoskeletal organization. Here, we examined...... the effect of direct modulation of actin microfilament dynamics on adipocyte differentiation. Stabilizing actin filaments in hMSCs by siRNA-mediated knock down of the two main actin depolymerizing factors (ADFs): Cofilin 1 (CFL1) and Destrin (DSTN) or treating the cells by Phalloidin reduced adipocyte...

  1. Superficial parotidectomy versus retrograde partial superficial parotidectomy in treating benign salivary gland tumor (pleomorphic adenoma).

    Science.gov (United States)

    Emodi, Omri; El-Naaj, Imad Abu; Gordin, Arye; Akrish, Sharon; Peled, Micha

    2010-09-01

    Of all benign salivary gland tumors of the parotid gland, pleomorphic adenoma (mixed tumor) is the most common. It accounts for 60% to 70% of all benign tumors of the parotid gland. This neoplasm arises in patients in the fourth to sixth decade of life, with a female predominance. The surgical excision of this lesion continues to be the subject of major debate. The goal is to avoid facial disability yet attain complete resection without perforation of the capsule/pseudocapsule. The purpose of our study is to compare 2 surgical techniques performed at the Ear, Nose, and Throat and Maxillofacial Departments, Rambam Medical Center, Haifa, Israel, and determine which is preferable in treating this lesion. We reviewed 48 patients who underwent excision of pleomorphic adenoma of the parotid gland between 1996 and 2005 at Rambam Medical Center: 18 were treated surgically with the classical superficial parotidectomy (SP) technique, using an anterograde approach, and 30 were treated with retrograde partial superficial parotidectomy (PSP). We compared the 2 surgical techniques in terms of surgical time, histopathologic size of the lesion, amount of excised healthy parotid tissue, histologic margin, and the preservation of the capsule/pseudocapsule. We also made clinical records of temporary or definitive injury to the facial nerve, which branches of the facial nerve were temporarily or definitively injured, the occurrence of Frey syndrome, esthetic satisfaction, and the amount of recurrence or infection after surgery. Of the 48 patients, 19 (39.6%) were male and 29 (60.4%) were female, with a mean age (+/- SD) of 43.8 +/- 16.97 years (median, 50 years; range, 12-79 years). We found a significant difference (P = .029) in mean surgical time (+/- SD): 171 +/- 49.7 minutes (median, 165 minutes) when performing the classical SP and 145 +/- 42.7 minutes (median, 130 minutes) when performing the retrograde PSP. Much more healthy parotid tissue was taken out with the classical

  2. Comparative histochemical study of Bowen’s disease and actinic keratosis: preserved normal basal cells in Bowen’s disease

    Directory of Open Access Journals (Sweden)

    H Ishida

    2009-12-01

    Full Text Available The degree of DNA-instability as revealed by immunohistochemical staining with anti-cytidine antibody after acid hydrolysis (DNA-instability test has been recently used as a marker of malignancy. This technique was applied to examine 17 skin tissue samples of Bowen’s disease, 47 of actinic keratosis, 15 of squamous cell carcinoma, 5 of seborrheic keratosis, and 10 of normal skin. All benign neoplastic cells of seborrheic keratosis and normal epidermal cells were negative. On the other hand, all cancer cells were positive with the DNA-instability test, indicating their malignancy, but all basal cells in Bowen’s disease were completely negative. Compatible with this result, the basal cells in Bowen’s disease were characteristically normal as evident in other histochemical examinations. Thus, they were negative with p53 immunohistochemistry, with normal signals of chromosome 17 in situ hybridisation and argyrophilic nucleolar organiser region, and showed slightly enhanced proliferative activity as revealed by proliferating cell nuclear antigen immunohistochemistry. Immunohistochemical staining with 34 ß E12 (monoclonal antibody against cytokeratins 1, 5, 10, and 14, which stains all normal epidermal keratinocytes including basal cells, showed that only the basal cells of Bowen’s disease stained strongly and homogeneously, while all cancer cells in the upper layers of Bowen’s disease and all layers of actinic keratosis were only sporadically or weakly stained. Staining with 34 ß B4 (monoclonal antibody against cytokeratin 1, which recognises the whole epidermis except for the basal layer in the normal epidermis, showed that the basal cells in the Bowen’s disease were completely negative, and lower layer cells in the actinic keratosis and upper layer cells in Bowen’s disease were only sporadically stained positive, although the superficial layer cells in actinic keratosis stained strongly and homogeneously. Our findings clearly

  3. Nanosecond electric pulses trigger actin responses in plant cells

    International Nuclear Information System (INIS)

    Berghoefer, Thomas; Eing, Christian; Flickinger, Bianca; Hohenberger, Petra; Wegner, Lars H.; Frey, Wolfgang; Nick, Peter

    2009-01-01

    We have analyzed the cellular effects of nanosecond pulsed electrical fields on plant cells using fluorescently tagged marker lines in the tobacco cell line BY-2 and confocal laser scanning microscopy. We observe a disintegration of the cytoskeleton in the cell cortex, followed by contraction of actin filaments towards the nucleus, and disintegration of the nuclear envelope. These responses are accompanied by irreversible permeabilization of the plasma membrane manifest as uptake of Trypan Blue. By pretreatment with the actin-stabilizing drug phalloidin, the detachment of transvacuolar actin from the cell periphery can be suppressed, and this treatment can also suppress the irreversible perforation of the plasma membrane. We discuss these findings in terms of a model, where nanosecond pulsed electric fields trigger actin responses that are key events in the plant-specific form of programmed cell death.

  4. Gamma Activation Analysis in the Havana Bay superficial sediments

    International Nuclear Information System (INIS)

    Lopez, N.; Gelen, A.; Diaz Riso, O.; Manso, M.V.; Simon, M.J.; Maslov, A.G.; Gustova, M.V.; Beltran, J.; Soto, J.

    2003-01-01

    A preliminary study of 26 elements of Havana Bay superficial sediments were made using Gamma Activation Analysis. Samples from five zones of Havana Bay were analyzed. The results show a close interrelation between the concentration levels of the studied elements and the contaminant sources

  5. Volumetric Modulated Arc Therapy (VMAT) Treatment Planning for Superficial Tumors

    International Nuclear Information System (INIS)

    Zacarias, Albert S.; Brown, Mellonie F.; Mills, Michael D.

    2010-01-01

    The physician's planning objective is often a uniform dose distribution throughout the planning target volume (PTV), including superficial PTVs on or near the surface of a patient's body. Varian's Eclipse treatment planning system uses a progressive resolution optimizer (PRO), version 8.2.23, for RapidArc dynamic multileaf collimator volumetric modulated arc therapy planning. Because the PRO is a fast optimizer, optimization convergence errors (OCEs) produce dose nonuniformity in the superficial area of the PTV. We present a postsurgical cranial case demonstrating the recursive method our clinic uses to produce RapidArc treatment plans. The initial RapidArc treatment plan generated using one 360 o arc resulted in substantial dose nonuniformity in the superficial section of the PTV. We demonstrate the use of multiple arcs to produce improved dose uniformity in this region. We also compare the results of this superficial dose compensation method to the results of a recursive method of dose correction that we developed in-house to correct optimization convergence errors in static intensity-modulated radiation therapy treatment plans. The results show that up to 4 arcs may be necessary to provide uniform dose to the surface of the PTV with the current version of the PRO.

  6. A method for determination of the superficial charge density

    International Nuclear Information System (INIS)

    Vila, F.

    1992-10-01

    In this article is presented a new methodism for determination of superficial charge density in nonconducting materials which is based in the combination of laboratory calibrated experiments in conducting surfaces with theoretical calculations for nonconducting surfaces. (author). 19 refs, 7 figs, 1 tab

  7. Superficial skin infections and the use of topical and systemic ...

    African Journals Online (AJOL)

    Superficial bacterial infections of the skin are very common. With the increasing burden of human immunodeficiency virus (HIV), this is likely to worsen. Examples of such infections include impetigo, erysipelas, cellulitis, ecthyma, furuncles, carbuncles and subcutaneous abscesses. Common causative organisms are ...

  8. Frey′s Syndrome as a Sequela of Superficial Parotidectomy

    Directory of Open Access Journals (Sweden)

    Rishi Kumar Bali

    2006-01-01

    The disorder is characterized by unilateral sweating and flushing of facial skin in the area of parotid gland occurring during meals. We present a case of a patient who developed symptoms of Frey Syndrome 4 months after undergoing superficial parotidectomy on left side.

  9. Diagnostic and therapeutic challenges in superficial CNS siderosis

    DEFF Research Database (Denmark)

    Kondziella, Daniel; Lindelof, M.; Haziri, Donika

    2015-01-01

    the challenges related to the diagnosis and treatment of superficial siderosis. RESULTS: A potential bleeding aetiology was identified in all patients, but removal of the offending bleeding source was achieved only in three (33%). Symptom progression was halted in just one patient (11%), which suggests...... neurotoxicity due to accumulating iron toxicity. FUNDING: not relevant. TRIAL REGISTRATION: not relevant....

  10. Superficial siderosis of the central nervous system | Govind | SA ...

    African Journals Online (AJOL)

    progression of clinical deterioration but halting the chronic subarachoid haemorrhage will reduce the iron load in the CSF and hence the cytotoxic risk. We present a case of superficial siderosis presenting with hearing loss and cerebellar signs highlighting the imaging findings. South African Journal of Radiology Vol.

  11. Treatment for superficial infusion thrombophlebitis of the upper extremity

    NARCIS (Netherlands)

    Di Nisio, Marcello; Peinemann, Frank; Porreca, Ettore; Rutjes, Anne W. S.

    2015-01-01

    Although superficial thrombophlebitis of the upper extremity represents a frequent complication of intravenous catheters inserted into the peripheral veins of the forearm or hand, no consensus exists on the optimal management of this condition in clinical practice. To summarise the evidence from

  12. Risk factors associated with acquiring superficial fungal infections in ...

    African Journals Online (AJOL)

    extensively documented. It is therefore imperative to identify the factors that predispose to the development of SFI so as to provide evidence-based and effective pre- ventive measures, thus reducing the prevalence and the attendant morbidity associated with superficial fungal in- fections and this was our aim in this present ...

  13. Bulkiness or aromatic nature of tyrosine-143 of actin is important for the weak binding between F-actin and myosin-ADP-phosphate

    Energy Technology Data Exchange (ETDEWEB)

    Gomibuchi, Yuki [Graduate School of Science and Engineering, Teikyo University, Toyosatodai 1-1, Utsunomiya 320-8551 (Japan); Uyeda, Taro Q.P. [Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, AIST Tsukuba Central 4, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562 (Japan); Wakabayashi, Takeyuki, E-mail: tw007@nasu.bio.teikyo-u.ac.jp [Graduate School of Science and Engineering, Teikyo University, Toyosatodai 1-1, Utsunomiya 320-8551 (Japan); Department of Judo Therapy, Faculty of Medical Technology, Teikyo University, Toyosatodai 1-1, Utsunomiya 320-8551 (Japan)

    2013-11-29

    Highlights: •The effect of mutation of Tyr143 that becomes more exposed on assembly was examined. •Mutation of tyrosine-143 of Dictyostelium actin changed actin polymerizability. •The bulkiness or aromatic nature of Tyr143 is important for the weak binding. •The weak interaction between myosin and actin strengthened by Tyr143Trp mutation. -- Abstract: Actin filaments (F-actin) interact with myosin and activate its ATPase to support force generation. By comparing crystal structures of G-actin and the quasi-atomic model of F-actin based on high-resolution cryo-electron microscopy, the tyrosine-143 was found to be exposed more than 60 Å{sup 2} to the solvent in F-actin. Because tyrosine-143 flanks the hydrophobic cleft near the hydrophobic helix that binds to myosin, the mutant actins, of which the tyrosine-143 was replaced with tryptophan, phenylalanine, or isoleucine, were generated using the Dictyostelium expression system. It polymerized significantly poorly when induced by NaCl, but almost normally by KCl. In the presence of phalloidin and KCl, the extents of the polymerization of all the mutant actins were comparable to that of the wild-type actin so that the actin-activated myosin ATPase activity could be reliably compared. The affinity of skeletal heavy meromyosin to F-actin and the maximum ATPase activity (V{sub max}) were estimated by a double reciprocal plot. The Tyr143Trp-actin showed the higher affinity (smaller K{sub app}) than that of the wild-type actin, with the V{sub max} being almost unchanged. The K{sub app} and V{sub max} of the Tyr143Phe-actin were similar to those of the wild-type actin. However, the activation by Tyr143Ile-actin was much smaller than the wild-type actin and the accurate determination of K{sub app} was difficult. Comparison of the myosin ATPase activated by the various mutant actins at the same concentration of F-actin showed that the extent of activation correlates well with the solvent-accessible surface areas (ASA

  14. The role of antihistamines in chronic actinic dermatitis treatment

    Directory of Open Access Journals (Sweden)

    E. V. Orlov

    2016-01-01

    Full Text Available Inveterate actinic dermatitis is an immunologically mediated photodermatosis characterized by itchy eczematous dermhelminthiasis exposed to sunlight. The disease proceeds in the same way as the atopic eczema or atopic dermatitis. The treatment of patients with inveterate actinic dermatitis is similar to the treatment of patients with atopic dermatitis and eczema. Administration of the modern antihistaminic preparation desloratadine (Aerius in the treatment has a positive effect on the skin process relief and on some cellular and humoral immunity factors.

  15. Actinic prurigo in Scandinavian adolescent successfully treated with cyclosporine A

    Directory of Open Access Journals (Sweden)

    Jan C. Sitek

    2017-06-01

    Full Text Available Actinic prurigo is a pruritic sun-induced dermatosis classified among the immunologically mediated photodermatoses. The disease is a well-known entity among Native Americans and in Central and South America, however rare in Caucasians with only a few reports from Australia, Britain and France. We report the first case of actinic prurigo in a Scandinavian patient, responding favorably to systemic treatment with cyclosporine A.

  16. Actinic Prurigo in Scandinavian Adolescent Successfully Treated with Cyclosporine A.

    Science.gov (United States)

    Sitek, Jan C

    2017-03-13

    Actinic prurigo is a pruritic sun-induced dermatosis classified among the immunologically mediated photodermatoses. The disease is a well-known entity among Native Americans and in Central and South America, however rare in Caucasians with only a few reports from Australia, Britain and France. We report the first case of actinic prurigo in a Scandinavian patient, responding favorably to systemic treatment with cyclosporine A.

  17. Amphidinolide H, a novel type of actin-stabilizing agent isolated from dinoflagellate

    International Nuclear Information System (INIS)

    Saito, Shin-ya; Feng Jue; Kira, Atsushi; Kobayashi, Jun'ichi; Ohizumi, Yasushi

    2004-01-01

    The effect of novel cytotoxic marine macrolide, amphidinolide H (Amp-H), on actin dynamics was investigated in vitro. Amp-H attenuated actin depolymerization induced by diluting F-actin. This effect remained after washing out of unbound Amp-H by filtration. In the presence of either Amp-H or phalloidin, lag phase, which is the rate-limiting step of actin polymerization, was shortened. Phalloidin decreased the polymerization-rate whereas Amp-H did not. Meanwhile, the effects of both compounds were the same when barbed end of actin was capped by cytochalasin D. Quartz crystal microbalance system revealed interaction of Amp-H with G-actin and F-actin. Amp-H also enhanced the binding of phalloidin to F-actin. We concluded that Amp-H stabilizes actin in a different manner from that of phalloidin and serves as a novel pharmacological tool for analyzing actin-mediated cell function

  18. Major Superficial White Matter Abnormalities in Huntington's Disease

    Science.gov (United States)

    Phillips, Owen R.; Joshi, Shantanu H.; Squitieri, Ferdinando; Sanchez-Castaneda, Cristina; Narr, Katherine; Shattuck, David W.; Caltagirone, Carlo; Sabatini, Umberto; Di Paola, Margherita

    2016-01-01

    Background: The late myelinating superficial white matter at the juncture of the cortical gray and white matter comprising the intracortical myelin and short-range association fibers has not received attention in Huntington's disease. It is an area of the brain that is late myelinating and is sensitive to both normal aging and neurodegenerative disease effects. Therefore, it may be sensitive to Huntington's disease processes. Methods: Structural MRI data from 25 Pre-symptomatic subjects, 24 Huntington's disease patients and 49 healthy controls was run through a cortical pattern-matching program. The surface corresponding to the white matter directly below the cortical gray matter was then extracted. Individual subject's Diffusion Tensor Imaging (DTI) data was aligned to their structural MRI data. Diffusivity values along the white matter surface were then sampled at each vertex point. DTI measures with high spatial resolution across the superficial white matter surface were then analyzed with the General Linear Model to test for the effects of disease. Results: There was an overall increase in the axial and radial diffusivity across much of the superficial white matter (p < 0.001) in Pre-symptomatic subjects compared to controls. In Huntington's disease patients increased diffusivity covered essentially the whole brain (p < 0.001). Changes are correlated with genotype (CAG repeat number) and disease burden (p < 0.001). Conclusions: This study showed broad abnormalities in superficial white matter even before symptoms are present in Huntington's disease. Since, the superficial white matter has a unique microstructure and function these abnormalities suggest it plays an important role in the disease. PMID:27242403

  19. Monoubiquitination Inhibits the Actin Bundling Activity of Fascin.

    Science.gov (United States)

    Lin, Shengchen; Lu, Shuang; Mulaj, Mentor; Fang, Bin; Keeley, Tyler; Wan, Lixin; Hao, Jihui; Muschol, Martin; Sun, Jianwei; Yang, Shengyu

    2016-12-30

    Fascin is an actin bundling protein that cross-links individual actin filaments into straight, compact, and stiff bundles, which are crucial for the formation of filopodia, stereocillia, and other finger-like membrane protrusions. The dysregulation of fascin has been implicated in cancer metastasis, hearing loss, and blindness. Here we identified monoubiquitination as a novel mechanism that regulates fascin bundling activity and dynamics. The monoubiquitination sites were identified to be Lys 247 and Lys 250 , two residues located in a positive charge patch at the actin binding site 2 of fascin. Using a chemical ubiquitination method, we synthesized chemically monoubiquitinated fascin and determined the effects of monoubiquitination on fascin bundling activity and dynamics. Our data demonstrated that monoubiquitination decreased the fascin bundling EC 50 , delayed the initiation of bundle assembly, and accelerated the disassembly of existing bundles. By analyzing the electrostatic properties on the solvent-accessible surface of fascin, we proposed that monoubiquitination introduced steric hindrance to interfere with the interaction between actin filaments and the positively charged patch at actin binding site 2. We also identified Smurf1 as a E3 ligase regulating the monoubiquitination of fascin. Our findings revealed a previously unidentified regulatory mechanism for fascin, which will have important implications for the understanding of actin bundle regulation under physiological and pathological conditions. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  20. Monoubiquitination Inhibits the Actin Bundling Activity of Fascin*

    Science.gov (United States)

    Lin, Shengchen; Lu, Shuang; Mulaj, Mentor; Fang, Bin; Keeley, Tyler; Wan, Lixin; Hao, Jihui; Muschol, Martin; Sun, Jianwei; Yang, Shengyu

    2016-01-01

    Fascin is an actin bundling protein that cross-links individual actin filaments into straight, compact, and stiff bundles, which are crucial for the formation of filopodia, stereocillia, and other finger-like membrane protrusions. The dysregulation of fascin has been implicated in cancer metastasis, hearing loss, and blindness. Here we identified monoubiquitination as a novel mechanism that regulates fascin bundling activity and dynamics. The monoubiquitination sites were identified to be Lys247 and Lys250, two residues located in a positive charge patch at the actin binding site 2 of fascin. Using a chemical ubiquitination method, we synthesized chemically monoubiquitinated fascin and determined the effects of monoubiquitination on fascin bundling activity and dynamics. Our data demonstrated that monoubiquitination decreased the fascin bundling EC50, delayed the initiation of bundle assembly, and accelerated the disassembly of existing bundles. By analyzing the electrostatic properties on the solvent-accessible surface of fascin, we proposed that monoubiquitination introduced steric hindrance to interfere with the interaction between actin filaments and the positively charged patch at actin binding site 2. We also identified Smurf1 as a E3 ligase regulating the monoubiquitination of fascin. Our findings revealed a previously unidentified regulatory mechanism for fascin, which will have important implications for the understanding of actin bundle regulation under physiological and pathological conditions. PMID:27879315

  1. Actin cytoskeleton modulates calcium signaling during maturation of starfish oocytes.

    Science.gov (United States)

    Kyozuka, Keiichiro; Chun, Jong T; Puppo, Agostina; Gragnaniello, Gianni; Garante, Ezio; Santella, Luigia

    2008-08-15

    Before successful fertilization can occur, oocytes must undergo meiotic maturation. In starfish, this can be achieved in vitro by applying 1-methyladenine (1-MA). The immediate response to 1-MA is the fast Ca2+ release in the cell cortex. Here, we show that this Ca2+ wave always initiates in the vegetal hemisphere and propagates through the cortex, which is the space immediately under the plasma membrane. We have observed that alteration of the cortical actin cytoskeleton by latrunculin-A and jasplakinolide can potently affect the Ca2+ waves triggered by 1-MA. This indicates that the cortical actin cytoskeleton modulates Ca2+ release during meiotic maturation. The Ca2+ wave was inhibited by the classical antagonists of the InsP(3)-linked Ca2+ signaling pathway, U73122 and heparin. To our surprise, however, these two inhibitors induced remarkable actin hyper-polymerization in the cell cortex, suggesting that their inhibitory effect on Ca2+ release may be attributed to the perturbation of the cortical actin cytoskeleton. In post-meiotic eggs, U73122 and jasplakinolide blocked the elevation of the vitelline layer by uncaged InsP(3), despite the massive release of Ca2+, implying that exocytosis of the cortical granules requires not only a Ca2+ rise, but also regulation of the cortical actin cytoskeleton. Our results suggest that the cortical actin cytoskeleton of starfish oocytes plays critical roles both in generating Ca2+ signals and in regulating cortical granule exocytosis.

  2. Prolactin promotes breast cancer cell migration through actin cytoskeleton remodeling

    Directory of Open Access Journals (Sweden)

    Priscilla Ludovico da Silva

    2015-12-01

    Full Text Available The role of prolactin on breast cancer development and progression is debated. Breast cancer progression largely depends on cell movement and on the ability to remodel the actin cytoskeleton. In this process, actin-binding proteins are requested to achieve fibrillar actin de-polymerization and relocation at the cell membrane. Kinases such as focal adhesion kinase (FAK are later required to form actin/vinculin-enriched structures called focal adhesion complexes, which mediate firm adhesion to the extracellular matrix. These controllers are regulated by c-Src, which forms multiprotein signaling complexes with membrane receptors and is regulated by a number of hormones, including prolactin. We here show that breast cancer cells exposed to prolactin display an elevated c-Src expression and phosphorylation. In parallel, increased moesin and FAK expression and phosphorylation are found. These molecular changes are associated to relocation to the plasma membrane of cytoskeletal actin fibers and to increased horizontal cell movement. In conclusion, prolactin regulates actin remodeling and enhances breast cancer cell movement. This finding broadens the understanding of prolactin actions on breast cancer cells, highlighting new pathways that may be relevant to on breast cancer progression.

  3. Functional characterisation of filamentous actin probe expression in neuronal cells.

    Directory of Open Access Journals (Sweden)

    Shrujna Patel

    Full Text Available Genetically encoded filamentous actin probes, Lifeact, Utrophin and F-tractin, are used as tools to label the actin cytoskeleton. Recent evidence in several different cell types indicates that these probes can cause changes in filamentous actin dynamics, altering cell morphology and function. Although these probes are commonly used to visualise actin dynamics in neurons, their effects on axonal and dendritic morphology has not been systematically characterised. In this study, we quantitatively analysed the effect of Lifeact, Utrophin and F-tractin on neuronal morphogenesis in primary hippocampal neurons. Our data show that the expression of actin-tracking probes significantly impacts on axonal and dendrite growth these neurons. Lifeact-GFP expression, under the control of a pBABE promoter, caused a significant decrease in total axon length, while another Lifeact-GFP expression, under the control of a CAG promoter, decreased the length and complexity of dendritic trees. Utr261-EGFP resulted in increased dendritic branching but Utr230-EGFP only accumulated in cell soma, without labelling any neurites. Lifeact-7-mEGFP and F-tractin-EGFP in a pEGFP-C1 vector, under the control of a CMV promoter, caused only minor changes in neuronal morphology as detected by Sholl analysis. The results of this study demonstrate the effects that filamentous actin tracking probes can have on the axonal and dendritic compartments of neuronal cells and emphasise the care that must be taken when interpreting data from experiments using these probes.

  4. Nuclear actin filaments recruit cofilin and actin-related protein 3, and their formation is connected with a mitotic block

    Czech Academy of Sciences Publication Activity Database

    Kalendová, Alžběta; Kalasová, Ilona; Yamazaki, S.; Uličná, Lívia; Harata, M.; Hozák, Pavel

    2014-01-01

    Roč. 142, č. 2 (2014), s. 139-152 ISSN 0948-6143 R&D Projects: GA ČR GAP305/11/2232; GA MŠk LD12063; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:68378050 Keywords : nuclear actin * transcription * mitosis * actin-related protein 3 * cofilin Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.927, year: 2013

  5. Probing GFP-actin diffusion in living cells using fluorescence correlation spectroscopy

    International Nuclear Information System (INIS)

    Engelke, Hanna; Heinrich, Doris; Rädler, Joachim O.

    2010-01-01

    The cytoskeleton of eukaryotic cells is continuously remodeled by polymerization and depolymerization of actin. Consequently, the relative content of polymerized filamentous actin (F-actin) and monomeric globular actin (G-actin) is subject to temporal and spatial fluctuations. Since fluorescence correlation spectroscopy (FCS) can measure the diffusion of fluorescently labeled actin it seems likely that FCS allows us to determine the dynamics and hence indirectly the structural properties of the cytoskeleton components with high spatial resolution. To this end we investigate the FCS signal of GFP-actin in living Dictyostelium discoideum cells and explore the inherent spatial and temporal signatures of the actin cytoskeleton. Using the free green fluorescent protein (GFP) as a reference, we find that actin diffusion inside cells is dominated by G-actin and slower than diffusion in diluted cell extract. The FCS signal in the dense cortical F-actin network near the cell membrane is probed using the cytoskeleton protein LIM and is found to be slower than cytosolic G-actin diffusion. Furthermore, we show that polymerization of the cytoskeleton induced by Jasplakinolide leads to a substantial decrease of G-actin diffusion. Pronounced fluctuations in the distribution of the FCS correlation curves can be induced by latrunculin, which is known to induce actin waves. Our work suggests that the FCS signal of GFP-actin in combination with scanning or spatial correlation techniques yield valuable information about the local dynamics and concomitant cytoskeletal properties

  6. A Novel Actinic Keratosis Field Assessment Scale for Grading Actinic Keratosis Disease Severity.

    Science.gov (United States)

    Dréno, Brigitte; Cerio, Rino; Dirschka, Thomas; Nart, Ignasi Figueras; Lear, John T; Peris, Ketty; de Casas, Andrés Ruiz; Kaleci, Shaniko; Pellacani, Giovanni

    2017-10-02

    Actinic keratosis (AK) lesions are surrounded by field cancerization (areas of subclinical, non-visible sun damage). Existing AK grading tools rely on AK counts, which are not reproducible. An Actinic Keratosis Field Assessment Scale (AK-FAS) for grading the severity of AK/field was developed. Standardized photographs of patients representing the full range of AK severity were collected. Six investigators independently rated each photograph according to 3 criteria: AK area (total skin area affected by AK lesions), hyperkeratosis and sun damage. Inter-rater reproducibility was good for all 3 criteria. Validation of the AK-FAS showed good reproducibility for AK area and hyperkeratosis, even for dermatologists untrained on use of the scale. In conclusion, the AK-FAS is objective, easy to use and implement, and reproducible. It incorporates assessment of the entire field affected by AK instead of relying on lesion counts. Use of the AK-FAS may standardize AK diagnosis, making it relevant to routine clinical practice.

  7. Efficacy of ingenol mebutate gel for actinic keratosis in patients treated by thiazide diuretics

    Directory of Open Access Journals (Sweden)

    Campione E

    2016-11-01

    Full Text Available Elena Campione,1 Monia Di Prete,1 Laura Diluvio,1 Luca Bianchi,1 Augusto Orlandi2 1Department of Dermatology, 2Department of Pathology, University of Rome Tor Vergata, Rome, Italy Abstract: Actinic keratosis (AK is considered as superficial squamous cell carcinoma. Chronic sun exposure plays a central role in its pathogenesis. In particular, ultraviolet B radiation causes direct damage to the DNA, producing pyrimidine dimers that suppress the protective role of p53. Other risk factors include advanced age, male sex, and fair skin type. Even some drugs used for treating blood hypertension, such as thiazide diuretics, can increase the risk of developing AK. Their photosensitizing action seems to be connected with reactive oxygen species production. We report our experience on ten patients affected by multiple AK, in therapy with thiazide diuretics, treated by ingenol mebutate gel. AK was clinically and dermoscopically evaluated at baseline and after 30 days from the beginning of the treatment. Moreover, patients were screened for vitamin D3 values and reported a general hypovitaminosis status. To our knowledge, we report for the first time the efficacy of ingenol mebutate gel in this group of patients, particularly at risk of developing AK. Keywords: anticancer agents, antitumor activity, cancer cells, skin precancerous lesion

  8. Actin depolymerization enhances adipogenic differentiation in human stromal stem cells

    Directory of Open Access Journals (Sweden)

    Li Chen

    2018-05-01

    Full Text Available Human stromal stem cells (hMSCs differentiate into adipocytes that play a role in skeletal tissue homeostasis and whole body energy metabolism. During adipocyte differentiation, hMSCs exhibit significant changes in cell morphology suggesting changes in cytoskeletal organization. Here, we examined the effect of direct modulation of actin microfilament dynamics on adipocyte differentiation. Stabilizing actin filaments in hMSCs by siRNA-mediated knock down of the two main actin depolymerizing factors (ADFs: Cofilin 1 (CFL1 and Destrin (DSTN or treating the cells by Phalloidin reduced adipocyte differentiation as evidenced by decreased number of mature adipocytes and decreased adipocyte specific gene expression (ADIPOQ, LPL, PPARG, FABP4. In contrast, disruption of actin cytoskeleton by Cytochalasin D enhanced adipocyte differentiation. Follow up studies revealed that the effects of CFL1 on adipocyte differentiation depended on the activity of LIM domain kinase 1 (LIMK1 which is the major upstream kinase of CFL1. Inhibiting LIMK by its specific chemical inhibitor LIMKi inhibited the phosphorylation of CFL1 and actin polymerization, and enhanced the adipocyte differentiation. Moreover, treating hMSCs by Cytochalasin D inhibited ERK and Smad2 signaling and this was associated with enhanced adipocyte differentiation. On the other hand, Phalloidin enhanced ERK and Smad2 signaling, but inhibited adipocyte differentiation which was rescued by ERK specific chemical inhibitor U0126. Our data provide a link between restructuring of hMSCs cytoskeleton and hMSCs lineage commitment and differentiation. Keywords: Actin cytoskeleton, Actin depolymerizing factors, Adipocyte differentiation, Human stromal stem cells

  9. Actin-cytoskeleton rearrangement modulates proton-induced uptake

    Energy Technology Data Exchange (ETDEWEB)

    Ben-Dov, Nadav [Department of Physiology and Pharmacology, Faculty of Medicine, Tel-Aviv University, 69978 Tel-Aviv (Israel); Korenstein, Rafi, E-mail: korens@post.tau.ac.il [Department of Physiology and Pharmacology, Faculty of Medicine, Tel-Aviv University, 69978 Tel-Aviv (Israel)

    2013-04-15

    Recently it has been shown that elevating proton concentration at the cell surface stimulates the formation of membrane invaginations and vesicles accompanied by an enhanced uptake of macromolecules. While the initial induction of inward membrane curvature was rationalized in terms of proton-based increase of charge asymmetry across the membrane, the mechanisms underlying vesicle formation and its scission are still unknown. In light of the critical role of actin in vesicle formation during endocytosis, the present study addresses the involvement of cytoskeletal actin in proton-induced uptake (PIU). The uptake of dextran-FITC is used as a measure for the factual fraction of inward invaginations that undergo scission from the cell's plasma membrane. Our findings show that the rate of PIU in suspended cells is constant, whereas the rate of PIU in adherent cells is gradually increased in time, saturating at the level possessed by suspended cells. This is consistent with pH induced gradual degradation of stress-fibers in adherent cells. Wortmannin and calyculin-A are able to elevate PIU by 25% in adherent cells but not in suspended cells, while cytochalasin-D, rapamycin and latrunculin-A elevate PIU both in adherent and suspended cells. However, extensive actin depolymerization by high concentrations of latrunculin-A is able to inhibit PIU. We conclude that proton-induced membrane vesiculation is restricted by the actin structural resistance to the plasma membrane bending. Nevertheless, a certain degree of cortical actin restructuring is required for the completion of the scission process. - Highlights: ► Acidification of cells' exterior enhances uptake of macromolecules by the cells. ► Disruption of actin stress fibers leads to enhancement of proton induced uptake. ► Extensive depolymerization of cellular actin attenuates proton-induced uptake.

  10. Enhanced gravitropism of roots with a disrupted cap actin cytoskeleton

    Science.gov (United States)

    Hou, Guichuan; Mohamalawari, Deepti R.; Blancaflor, Elison B.

    2003-01-01

    The actin cytoskeleton has been proposed to be a major player in plant gravitropism. However, understanding the role of actin in this process is far from complete. To address this problem, we conducted an analysis of the effect of Latrunculin B (Lat B), a potent actin-disrupting drug, on root gravitropism using various parameters that included detailed curvature kinetics, estimation of gravitropic sensitivity, and monitoring of curvature development after extended clinorotation. Lat B treatment resulted in a promotion of root curvature after a 90 degrees reorientation in three plant species tested. More significantly, the sensitivity of maize (Zea mays) roots to gravity was enhanced after actin disruption, as determined from a comparison of presentation time of Lat B-treated versus untreated roots. A short 10-min gravistimulus followed by extended rotation on a 1-rpm clinostat resulted in extensive gravitropic responses, manifested as curvature that often exceeded 90 degrees. Application of Lat B to the cap or elongation zone of maize roots resulted in the disruption of the actin cytoskeleton, which was confined to the area of localized Lat B application. Only roots with Lat B applied to the cap displayed the strong curvature responses after extended clinorotation. Our study demonstrates that disrupting the actin cytoskeleton in the cap leads to the persistence of a signal established by a previous gravistimulus. Therefore, actin could function in root gravitropism by providing a mechanism to regulate the proliferation of a gravitropic signal originating from the cap to allow the root to attain its correct orientation or set point angle.

  11. Superficial evolution and compacting aptitude of uranium dioxide powders

    International Nuclear Information System (INIS)

    Danroc, J.

    1982-04-01

    Long term storage of UO 2 powder improves slightly shaping and solidity of compacted powder. The aim of this work is the study of material evolution and the increase of this evolution rate for application to industrial fabrication. Aging in wet air at different temperatures is examined. Evolution of texture and superficial composition is followed. Below 80 0 C UO 3 , 2H 2 O is formed at crystal surface and thermal decomposition gives different hydrates. Kinetics of the transformation is studied. Oxidohydratation in liquid phase is rapid with hydrogen peroxide. The aged or treated material is compacted and mechanical behaviour is examined. Improvement is explained by inter-layer water molecule of the superficial hydrate giving lubricant and pseudo-plastic properties [fr

  12. Effects of superficial gas velocity and fluid property on the ...

    African Journals Online (AJOL)

    In the present study, the influence of superficial gas velocity and fluid properties on gas holdup and liquid circulation velocity in a three-phase external loop airlift column using polystyrene (0.0036 m diameter and 1025.55 kg/m3 density) and nylon-6 (0.0035 m diameter and 1084.24 kg/m3 density) particles with aqueous ...

  13. Isolated core vs. superficial cooling effects on virtual maze navigation.

    Science.gov (United States)

    Payne, Jennifer; Cheung, Stephen S

    2007-07-01

    Cold impairs cognitive performance and is a common occurrence in many survival situations. Altered behavior patterns due to impaired navigation abilities in cold environments are potential problems in lost-person situations. We investigated the separate effects of low core temperature and superficial cooling on a spatially demanding virtual navigation task. There were 12 healthy men who were passively cooled via 15 degrees C water immersion to a core temperature of 36.0 degrees C, then transferred to a warm (40 degrees C) water bath to eliminate superficial shivering while completing a series of 20 virtual computer mazes. In a control condition, subjects rested in a thermoneutral (approximately 35 degrees C) bath for a time-matched period before being transferred to a warm bath for testing. Superficial cooling and distraction were achieved by whole-body immersion in 35 degree water for a time-matched period, followed by lower leg immersion in 10 degree C water for the duration of the navigational tests. Mean completion time and mean error scores for the mazes were not significantly different (p > 0.05) across the core cooling (16.59 +/- 11.54 s, 0.91 +/- 1.86 errors), control (15.40 +/- 8.85 s, 0.82 +/- 1.76 errors), and superficial cooling (15.19 +/- 7.80 s, 0.77 +/- 1.40 errors) conditions. Separately reducing core temperature or increasing cold sensation in the lower extremities did not influence performance on virtual computer mazes, suggesting that navigation is more resistive to cooling than other, simpler cognitive tasks. Further research is warranted to explore navigational ability at progressively lower core and skin temperatures, and in different populations.

  14. Superficial or deep implantation of motor nerve after denervation: an experimental study--superficial or deep implantation of motor nerve.

    Science.gov (United States)

    Askar, Ibrahím; Sabuncuoglu, Bízden Tavíl

    2002-01-01

    Neurorraphy, conventional nerve grafting technique, and artificial nerve conduits are not enough for repair in severe injuries of peripheral nerves, especially when there is separation of motor nerve from muscle tissue. In these nerve injuries, reinnervation is indicated for neurotization. The distal end of a peripheral nerve is divided into fascicles and implanted into the aneural zone of target muscle tissue. It is not known how deeply fascicles should be implanted into muscle tissue. A comparative study of superficial and deep implantation of separated motor nerve into muscle tissue is presented in the gastrocnemius muscle of rabbits. In this experimental study, 30 white New Zealand rabbits were used and divided into 3 groups of 10 rabbits each. In the first group (controls, group I), only surgical exposure of the gastrocnemius muscle and motor nerve (tibial nerve) was done without any injury to nerves. In the superficial implantation group (group II), tibial nerves were separated and divided into their own fascicles. These fascicles were implanted superficially into the lateral head of gastrocnemius muscle-aneural zone. In the deep implantation group (group III), the tibial nerves were separated and divided into their own fascicles. These fascicles were implanted around the center of the muscle mass, into the lateral head of the gastrocnemius muscle-aneural zone. Six months later, histopathological changes and functional recovery of the gastrocnemius muscle were investigated. Both experimental groups had less muscular weight than in the control group. It was found that functional recovery was achieved in both experimental groups, and was better in the superficial implantation group than the deep implantation group. EMG recordings revealed that polyphasic and late potentials were frequently seen in both experimental groups. Degeneration and regeneration of myofibrils were observed in both experimental groups. New motor end-plates were formed in a scattered

  15. Automated detection of actinic keratoses in clinical photographs.

    Science.gov (United States)

    Hames, Samuel C; Sinnya, Sudipta; Tan, Jean-Marie; Morze, Conrad; Sahebian, Azadeh; Soyer, H Peter; Prow, Tarl W

    2015-01-01

    Clinical diagnosis of actinic keratosis is known to have intra- and inter-observer variability, and there is currently no non-invasive and objective measure to diagnose these lesions. The aim of this pilot study was to determine if automatically detecting and circumscribing actinic keratoses in clinical photographs is feasible. Photographs of the face and dorsal forearms were acquired in 20 volunteers from two groups: the first with at least on actinic keratosis present on the face and each arm, the second with no actinic keratoses. The photographs were automatically analysed using colour space transforms and morphological features to detect erythema. The automated output was compared with a senior consultant dermatologist's assessment of the photographs, including the intra-observer variability. Performance was assessed by the correlation between total lesions detected by automated method and dermatologist, and whether the individual lesions detected were in the same location as the dermatologist identified lesions. Additionally, the ability to limit false positives was assessed by automatic assessment of the photographs from the no actinic keratosis group in comparison to the high actinic keratosis group. The correlation between the automatic and dermatologist counts was 0.62 on the face and 0.51 on the arms, compared to the dermatologist's intra-observer variation of 0.83 and 0.93 for the same. Sensitivity of automatic detection was 39.5% on the face, 53.1% on the arms. Positive predictive values were 13.9% on the face and 39.8% on the arms. Significantly more lesions (p<0.0001) were detected in the high actinic keratosis group compared to the no actinic keratosis group. The proposed method was inferior to assessment by the dermatologist in terms of sensitivity and positive predictive value. However, this pilot study used only a single simple feature and was still able to achieve sensitivity of detection of 53.1% on the arms.This suggests that image analysis is

  16. Interdigital foot infections: Corynebacterium minutissimum and agents of superficial mycoses

    Directory of Open Access Journals (Sweden)

    Fatma Mutlu Sariguzel

    2014-09-01

    Full Text Available Interdigital foot infections are mostly caused initially by dermatophytes, yeasts and less frequently by bacteria. Erythrasma caused by Corynebacterium minutissimum can be confused with superficial mycoses. The aim of the study was to determine the prevalence of the etiologic agents of superficial mycoses and the frequency of Corynebacterium minutissimum in interdigital foot infections. All the samples obtained from the 121 patients with interdigital foot infections were examined directly with the use of 20% potassium hydroxide mounts and Gram stain under the microscope and cultured on Sabouraud's dextrose agar plates. In identification of superficial mycoses, the rate was found to be 14% with the cultural method and 14% with direct microscopic examination. Using a combination of direct microscopic examination and culture, a 33.8% ratio was achieved. In the culture of these samples, the most isolated factor was Trichophyton rubrum (33.7%. In 24 of the patients (19.8% Corynebacterium minutissimum was detected by Gram staining, in 6 of these patients Trichophyton rubrum was found, Trichophyton mentagrophytes was found in 2 and Trichosporon spp. was found in 1. The examination of interdigital foot lesions in the laboratory, the coexistence of erythrasma with dermatophytes and yeast should be considered.

  17. Superficies in the form of the right to superpose

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    Simona CHIRICĂ

    2015-06-01

    Full Text Available The purpose of this paper is to present the current legal framework related to the superficies right in the form of the right to superpose, and especially to draw the attention and put certain question marks regarding the actuality or even the urgency of the need for regulation regarding the right to superpose. First, as a preliminary aspect, in order to emphasize the historical evolution of the superficies right, we will briefly present the development of this concept starting from the Roman law up to the present date. Second, by analysing the relevant legislation, the doctrine and the jurisprudence, the authors set themselves to present the main methods for constituting the superficies right. Third, the characteristics related to the right to superpose will be correlatively laid out. Fourth, the possibility to obtain a building permit on the basis of the right to superpose will also be analysed. Fifth, the recently entered-into-force legislative framework regarding the registration of the right to superpose and of the building thus erected is presented. Last but not least, the conclusions of this paper are presented, highlighting the necessity for more clearly defined rules regulating the legal status of the right to superpose, in order to avoid any confusions and inconsistencies in practice.

  18. Incorporation of β-actin loading control into zymography.

    Science.gov (United States)

    Govindasamy, Natasha; Yan, MengJie; Jurasz, Paul

    2016-11-01

    Gelatin zymography and immunoblot are widely used gel electrophoresis techniques to study matrix metalloproteinases-2 and -9. Each method has its advantages and disadvantages. Zymography is exquisitely sensitive but offers no loading control to ensure equal sample loading. Immunoblot is a 100-1000-fold less sensitive, but allows for the probing of a sample loading control such as β-actin to ensure accurate protein loading. In this report, we describe two simple protocols that combine gelatin zymography to study MMP-2 and -9 levels with an in-gel β-actin immunoblot loading control, thus combining sensitivity and accuracy in a single assay. The protocols incorporate the loading of molecular weight markers to demarcate MMP-2/-9 from the β-actin. The first protocol utilizes the overlay of a 10% zymography gel over a 5% Tris-Glycine separating gel from which the β-actin is transferred. The second protocol involves the direct transfer of the β-actin from a single 10% zymography gel.

  19. Hippocampal Dendritic Spines Are Segregated Depending on Their Actin Polymerization.

    Science.gov (United States)

    Domínguez-Iturza, Nuria; Calvo, María; Benoist, Marion; Esteban, José Antonio; Morales, Miguel

    2016-01-01

    Dendritic spines are mushroom-shaped protrusions of the postsynaptic membrane. Spines receive the majority of glutamatergic synaptic inputs. Their morphology, dynamics, and density have been related to synaptic plasticity and learning. The main determinant of spine shape is filamentous actin. Using FRAP, we have reexamined the actin dynamics of individual spines from pyramidal hippocampal neurons, both in cultures and in hippocampal organotypic slices. Our results indicate that, in cultures, the actin mobile fraction is independently regulated at the individual spine level, and mobile fraction values do not correlate with either age or distance from the soma. The most significant factor regulating actin mobile fraction was the presence of astrocytes in the culture substrate. Spines from neurons growing in the virtual absence of astrocytes have a more stable actin cytoskeleton, while spines from neurons growing in close contact with astrocytes show a more dynamic cytoskeleton. According to their recovery time, spines were distributed into two populations with slower and faster recovery times, while spines from slice cultures were grouped into one population. Finally, employing fast lineal acquisition protocols, we confirmed the existence of loci with high polymerization rates within the spine.

  20. All-Round Manipulation of the Actin Cytoskeleton by HIV.

    Science.gov (United States)

    Ospina Stella, Alberto; Turville, Stuart

    2018-02-05

    While significant progress has been made in terms of human immunodeficiency virus (HIV) therapy, treatment does not represent a cure and remains inaccessible to many people living with HIV. Continued mechanistic research into the viral life cycle and its intersection with many aspects of cellular biology are not only fundamental in the continued fight against HIV, but also provide many key observations of the workings of our immune system. Decades of HIV research have testified to the integral role of the actin cytoskeleton in both establishing and spreading the infection. Here, we review how the virus uses different strategies to manipulate cellular actin networks and increase the efficiency of various stages of its life cycle. While some HIV proteins seem able to bind to actin filaments directly, subversion of the cytoskeleton occurs indirectly by exploiting the power of actin regulatory proteins, which are corrupted at multiple levels. Furthermore, this manipulation is not restricted to a discrete class of proteins, but rather extends throughout all layers of the cytoskeleton. We discuss prominent examples of actin regulators that are exploited, neutralized or hijacked by the virus, and address how their coordinated deregulation can lead to changes in cellular behavior that promote viral spreading.

  1. Actin depolymerization enhances adipogenic differentiation in human stromal stem cells.

    Science.gov (United States)

    Chen, Li; Hu, Huimin; Qiu, Weimin; Shi, Kaikai; Kassem, Moustapha

    2018-05-01

    Human stromal stem cells (hMSCs) differentiate into adipocytes that play a role in skeletal tissue homeostasis and whole body energy metabolism. During adipocyte differentiation, hMSCs exhibit significant changes in cell morphology suggesting changes in cytoskeletal organization. Here, we examined the effect of direct modulation of actin microfilament dynamics on adipocyte differentiation. Stabilizing actin filaments in hMSCs by siRNA-mediated knock down of the two main actin depolymerizing factors (ADFs): Cofilin 1 (CFL1) and Destrin (DSTN) or treating the cells by Phalloidin reduced adipocyte differentiation as evidenced by decreased number of mature adipocytes and decreased adipocyte specific gene expression (ADIPOQ, LPL, PPARG, FABP4). In contrast, disruption of actin cytoskeleton by Cytochalasin D enhanced adipocyte differentiation. Follow up studies revealed that the effects of CFL1 on adipocyte differentiation depended on the activity of LIM domain kinase 1 (LIMK1) which is the major upstream kinase of CFL1. Inhibiting LIMK by its specific chemical inhibitor LIMKi inhibited the phosphorylation of CFL1 and actin polymerization, and enhanced the adipocyte differentiation. Moreover, treating hMSCs by Cytochalasin D inhibited ERK and Smad2 signaling and this was associated with enhanced adipocyte differentiation. On the other hand, Phalloidin enhanced ERK and Smad2 signaling, but inhibited adipocyte differentiation which was rescued by ERK specific chemical inhibitor U0126. Our data provide a link between restructuring of hMSCs cytoskeleton and hMSCs lineage commitment and differentiation. Copyright © 2018 Elsevier B.V. All rights reserved.

  2. Multiple roles for the actin cytoskeleton during regulated exocytosis

    Science.gov (United States)

    Porat-Shliom, Natalie; Milberg, Oleg; Masedunskas, Andrius; Weigert, Roberto

    2014-01-01

    Regulated exocytosis is the main mechanism utilized by specialized secretory cells to deliver molecules to the cell surface by virtue of membranous containers (i.e. secretory vesicles). The process involves a series of highly coordinated and sequential steps, which include the biogenesis of the vesicles, their delivery to the cell periphery, their fusion with the plasma membrane and the release of their content into the extracellular space. Each of these steps is regulated by the actin cytoskeleton. In this review, we summarize the current knowledge regarding the involvement of actin and its associated molecules during each of the exocytic steps in vertebrates, and suggest that the overall role of the actin cytoskeleton during regulated exocytosis is linked to the architecture and the physiology of the secretory cells under examination. Specifically, in neurons, neuroendocrine, endocrine, and hematopoietic cells, which contain small secretory vesicles that undergo rapid exocytosis (on the order of milliseconds), the actin cytoskeleton plays a role in pre-fusion events, where it acts primarily as a functional barrier and facilitates docking. In exocrine and other secretory cells, which contain large secretory vesicles that undergo slow exocytosis (seconds to minutes), the actin cytoskeleton plays a role in post-fusion events, where it regulates the dynamics of the fusion pore, facilitates the integration of the vesicles into the plasma membrane, provides structural support, and promotes the expulsion of large cargo molecules. PMID:22986507

  3. Addition of electrophilic lipids to actin alters filament structure

    International Nuclear Information System (INIS)

    Gayarre, Javier; Sanchez, David; Sanchez-Gomez, Francisco J.; Terron, Maria C.; Llorca, Oscar; Perez-Sala, Dolores

    2006-01-01

    Pathophysiological processes associated with oxidative stress lead to the generation of reactive lipid species. Among them, lipids bearing unsaturated aldehyde or ketone moieties can form covalent adducts with cysteine residues and modulate protein function. Through proteomic techniques we have identified actin as a target for the addition of biotinylated analogs of the cyclopentenone prostaglandins 15-deoxy-Δ 12,14 -PGJ 2 (15d-PGJ 2 ) and PGA 1 in NIH-3T3 fibroblasts. This modification could take place in vitro and mapped to the protein C-terminal end. Other electrophilic lipids, like the isoprostane 8-iso-PGA 1 and 4-hydroxy-2-nonenal, also bound to actin. The C-terminal region of actin is important for monomer-monomer interactions and polymerization. Electron microscopy showed that actin treated with 15d-PGJ 2 or 4-hydroxy-2-nonenal formed filaments which were less abundant and displayed shorter length and altered structure. Streptavidin-gold staining allowed mapping of biotinylated 15d-PGJ 2 at sites of filament disruption. These results shed light on the structural implications of actin modification by lipid electrophiles

  4. Spatially restricted actin-regulatory signaling contributes to synapse morphology

    Science.gov (United States)

    Nicholson, Daniel A.; Cahill, Michael E.; Tulisiak, Christopher T.; Geinisman, Yuri; Penzes, Peter

    2012-01-01

    The actin cytoskeleton in dendritic spines is organized into microdomains, but how signaling molecules that regulate actin are spatially governed is incompletely understood. Here we examine how the localization of the RacGEF kalirin-7, a well-characterized regulator of actin in spines, varies as a function of postsynaptic density (PSD) area and spine volume. Using serial section electron microscopy (EM), we find that extrasynaptic, but not synaptic, expression of kalirin-7 varies directly with synapse size and spine volume. Moreover, we find that overall expression levels of kalirin-7 differ in spines bearing perforated and non-perforated synapses, due primarily to extrasynaptic pools of kalirin-7 expression in the former. Overall, our findings indicate that kalirin-7 is differentially compartmentalized in spines as a function of both synapse morphology and spine size. PMID:22458534

  5. Formation of actin networks in microfluidic concentration gradients

    Directory of Open Access Journals (Sweden)

    Natalja eStrelnikova

    2016-05-01

    Full Text Available The physical properties of cytoskeletal networks are contributors in a number of mechanical responses of cells including cellular deformation and locomotion, and are crucial for the proper action of living cells. Local chemical gradients modulate cytoskeletal functionality including the interactions of the cytoskeleton with other cellular components. Actin is a major constituent of the cytoskeleton. Introducing a microfluidic-based platform, we explored the impact of concentration gradients on the formation and structural properties of actin networks. Microfluidics-controlled flow-free steady state experimental conditions allow for the generation of chemical gradients of different profiles, such as linear or step-like. We discovered specific features of actin networks emerging in defined gradients. In particular, we analyzed the effects of spatial conditions on network properties, bending rigidities of network links, and the network elasticity.

  6. Actin and Arp2/3 localize at the centrosome of interphase cells

    Energy Technology Data Exchange (ETDEWEB)

    Hubert, Thomas; Vandekerckhove, Joel; Gettemans, Jan, E-mail: jan.gettemans@vib-ugent.be

    2011-01-07

    Research highlights: {yields} Actin was detected at the centrosome with the anti-actin antibody 1C7 that recognizes antiparallel ('lower dimer') actin dimers. {yields} Centrosomal actin was found in interphase but not mitotic MDA-MB-231 cells. {yields} Neither the anti-actin antibody C4 that binds to globular, monomer actin, nor the anti-actin antibody 2G2 that recognizes the nuclear conformation of actin detect actin at the centrosome. {yields} The Arp2/3 complex transiently localizes at the pericentriolar matrix but not at the centrioles of interphase HEK 293T cells. -- Abstract: Although many actin binding proteins such as cortactin and the Arp2/3 activator WASH localize at the centrosome, the presence and conformation of actin at the centrosome has remained elusive. Here, we report the localization of actin at the centrosome in interphase but not in mitotic MDA-MB-231 cells. Centrosomal actin was detected with the anti-actin antibody 1C7 that recognizes antiparallel ('lower dimer') actin dimers. In addition, we report the transient presence of the Arp2/3 complex at the pericentriolar matrix but not at the centrioles of interphase HEK 293T cells. Overexpression of an Arp2/3 component resulted in expansion of the pericentriolar matrix and selective accumulation of the Arp2/3 component in the pericentriolar matrix. Altogether, we hypothesize that the centrosome transiently recruits Arp2/3 to perform processes such as centrosome separation prior to mitotic entry, whereas the observed constitutive centrosomal actin staining in interphase cells reinforces the current model of actin-based centrosome reorientation toward the leading edge in migrating cells.

  7. Actin and Arp2/3 localize at the centrosome of interphase cells

    International Nuclear Information System (INIS)

    Hubert, Thomas; Vandekerckhove, Joel; Gettemans, Jan

    2011-01-01

    Research highlights: → Actin was detected at the centrosome with the anti-actin antibody 1C7 that recognizes antiparallel ('lower dimer') actin dimers. → Centrosomal actin was found in interphase but not mitotic MDA-MB-231 cells. → Neither the anti-actin antibody C4 that binds to globular, monomer actin, nor the anti-actin antibody 2G2 that recognizes the nuclear conformation of actin detect actin at the centrosome. → The Arp2/3 complex transiently localizes at the pericentriolar matrix but not at the centrioles of interphase HEK 293T cells. -- Abstract: Although many actin binding proteins such as cortactin and the Arp2/3 activator WASH localize at the centrosome, the presence and conformation of actin at the centrosome has remained elusive. Here, we report the localization of actin at the centrosome in interphase but not in mitotic MDA-MB-231 cells. Centrosomal actin was detected with the anti-actin antibody 1C7 that recognizes antiparallel ('lower dimer') actin dimers. In addition, we report the transient presence of the Arp2/3 complex at the pericentriolar matrix but not at the centrioles of interphase HEK 293T cells. Overexpression of an Arp2/3 component resulted in expansion of the pericentriolar matrix and selective accumulation of the Arp2/3 component in the pericentriolar matrix. Altogether, we hypothesize that the centrosome transiently recruits Arp2/3 to perform processes such as centrosome separation prior to mitotic entry, whereas the observed constitutive centrosomal actin staining in interphase cells reinforces the current model of actin-based centrosome reorientation toward the leading edge in migrating cells.

  8. Hypertrophic stimulation increases beta-actin dynamics in adult feline cardiomyocytes.

    Directory of Open Access Journals (Sweden)

    Sundaravadivel Balasubramanian

    2010-07-01

    Full Text Available The myocardium responds to hemodynamic stress through cellular growth and organ hypertrophy. The impact of cytoskeletal elements on this process, however, is not fully understood. While alpha-actin in cardiomyocytes governs muscle contraction in combination with the myosin motor, the exact role of beta-actin has not been established. We hypothesized that in adult cardiomyocytes, as in non-myocytes, beta-actin can facilitate cytoskeletal rearrangement within cytoskeletal structures such as Z-discs. Using a feline right ventricular pressure overload (RVPO model, we measured the level and distribution of beta-actin in normal and pressure overloaded myocardium. Resulting data demonstrated enriched levels of beta-actin and enhanced translocation to the Triton-insoluble cytoskeletal and membrane skeletal complexes. In addition, RVPO in vivo and in vitro hypertrophic stimulation with endothelin (ET or insulin in isolated adult cardiomyocytes enhanced the content of polymerized fraction (F-actin of beta-actin. To determine the localization and dynamics of beta-actin, we adenovirally expressed GFP-tagged beta-actin in isolated adult cardiomyocytes. The ectopically expressed beta-actin-GFP localized to the Z-discs, costameres, and cell termini. Fluorescence recovery after photobleaching (FRAP measurements of beta-actin dynamics revealed that beta-actin at the Z-discs is constantly being exchanged with beta-actin from cytoplasmic pools and that this exchange is faster upon hypertrophic stimulation with ET or insulin. In addition, in electrically stimulated isolated adult cardiomyocytes, while beta-actin overexpression improved cardiomyocyte contractility, immunoneutralization of beta-actin resulted in a reduced contractility suggesting that beta-actin could be important for the contractile function of adult cardiomyocytes. These studies demonstrate the presence and dynamics of beta-actin in the adult cardiomyocyte and reinforce its usefulness in measuring

  9. How capping protein enhances actin filament growth and nucleation on biomimetic beads.

    Science.gov (United States)

    Wang, Ruizhe; Carlsson, Anders E

    2015-11-25

    Capping protein (CP), which caps the growing ends of actin filaments, accelerates actin-based motility. Recent experiments on biomimetic beads have shown that CP also enhances the rate of actin filament nucleation. Proposed explanations for these phenomena include (i) the actin funneling hypothesis (AFH), in which the presence of CP increases the free-actin concentration, and (ii) the monomer gating model, in which CP binding to actin filament barbed ends makes more monomers available for filament nucleation. To establish how CP increases the rates of filament elongation and nucleation on biomimetic beads, we perform a quantitative modeling analysis of actin polymerization, using rate equations that include actin filament nucleation, polymerization and capping, as modified by monomer depletion near the surface of the bead. With one adjustable parameter, our simulation results match previously measured time courses of polymerized actin and filament number. The results support a version of the AFH where CP increases the local actin monomer concentration at the bead surface, but leaves the global free-actin concentration nearly constant. Because the rate of filament nucleation increases with the monomer concentration, the increased local monomer concentration enhances actin filament nucleation. We derive a closed-form formula for the characteristic CP concentration where the local free-actin concentration reaches half the bulk value, and find it to be comparable to the global Arp2/3 complex concentration. We also propose an experimental protocol for distinguishing branching nucleation of filaments from spontaneous nucleation.

  10. Src kinases regulate de novo actin polymerization during exocytosis in neuroendocrine chromaffin cells.

    Directory of Open Access Journals (Sweden)

    María José Olivares

    Full Text Available The cortical actin network is dynamically rearranged during secretory processes. Nevertheless, it is unclear how de novo actin polymerization and the disruption of the preexisting actin network control transmitter release. Here we show that in bovine adrenal chromaffin cells, both formation of new actin filaments and disruption of the preexisting cortical actin network are induced by Ca2+ concentrations that trigger exocytosis. These two processes appear to regulate different stages of exocytosis; whereas the inhibition of actin polymerization with the N-WASP inhibitor wiskostatin restricts fusion pore expansion, thus limiting the release of transmitters, the disruption of the cortical actin network with cytochalasin D increases the amount of transmitter released per event. Further, the Src kinase inhibitor PP2, and cSrc SH2 and SH3 domains also suppress Ca2+-dependent actin polymerization, and slow down fusion pore expansion without disturbing the cortical F-actin organization. Finally, the isolated SH3 domain of c-Src prevents both the disruption of the actin network and the increase in the quantal release induced by cytochalasin D. These findings support a model where a rise in the cytosolic Ca2+ triggers actin polymerization through a mechanism that involves Src kinases. The newly formed actin filaments would speed up the expansion of the initial fusion pore, whereas the preexisting actin network might control a different step of the exocytosis process.

  11. Two Functionally Distinct Sources of Actin Monomers Supply the Leading Edge of Lamellipodia

    Science.gov (United States)

    Vitriol, Eric A.; McMillen, Laura M.; Kapustina, Maryna; Gomez, Shawn M.; Vavylonis, Dimitrios; Zheng, James Q.

    2015-01-01

    Summary Lamellipodia, the sheet-like protrusions of motile cells, consist of networks of actin filaments (F-actin) regulated by the ordered assembly from and disassembly into actin monomers (G-actin). Traditionally, G-actin is thought to exist as a homogeneous pool. Here, we show that there are two functionally and molecularly distinct sources of G-actin that supply lamellipodial actin networks. G-actin originating from the cytosolic pool requires the monomer binding protein thymosin β4 (Tβ4) for optimal leading edge localization, is targeted to formins, and is responsible for creating an elevated G/F-actin ratio that promotes membrane protrusion. The second source of G-actin comes from recycled lamellipodia F-actin. Recycling occurs independently of Tβ4 and appears to regulate lamellipodia homeostasis. Tβ4-bound G-actin specifically localizes to the leading edge because it doesn’t interact with Arp2/3-mediated polymerization sites found throughout the lamellipodia. These findings demonstrate that actin networks can be constructed from multiple sources of monomers with discrete spatiotemporal functions. PMID:25865895

  12. Modelling phagosomal lipid networks that regulate actin assembly

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    Schwarz Roland

    2008-12-01

    Full Text Available Abstract Background When purified phagosomes are incubated in the presence of actin under appropriate conditions, microfilaments start growing from the membrane in a process that is affected by ATP and the lipid composition of the membrane. Isolated phagosomes are metabolically active organelles that contain enzymes and metabolites necessary for lipid interconversion. Hence, addition of ATP, lipids, and actin to the system alter the steady-state composition of the phagosomal membrane at the same time that the actin nucleation is initiated. Our aim was to model all these processes in parallel. Results We compiled detailed experimental data on the effects of different lipids and ATP on actin nucleation and we investigated experimentally lipid interconversion and ATP metabolism in phagosomes by using suitable radioactive compounds. In a first step, a complex lipid network interconnected by chemical reactions catalyzed by known enzymes was modelled in COPASI (Complex Pathway Simulator. However, several lines of experimental evidence indicated that only the phosphatidylinositol branch of the network was active, an observation that dramatically reduced the number of parameters in the model. The results also indicated that a lipid network-independent ATP-consuming activity should be included in the model. When this activity was introduced, the set of differential equations satisfactorily reproduced the experimental data. On the other hand, a molecular mechanism connecting membrane lipids, ATP, and the actin nucleation process is still missing. We therefore adopted a phenomenological (black-box approach to represent the empirical observations. We proposed that lipids and ATP influence the dynamic interconversion between active and inactive actin nucleation sites. With this simple model, all the experimental data were satisfactorily fitted with a single positive parameter per lipid and ATP. Conclusion By establishing an active 'dialogue' between an

  13. Photodynamic therapy for actinic keratosis in organ transplant patients

    DEFF Research Database (Denmark)

    Basset-Seguin, N; Baumann Conzett, K; Gerritsen, M J P

    2013-01-01

    Background The incidence of actinic keratoses (AK) and non-melanoma skin cancer (NMSC) in organ transplant recipients (OTRs) is significantly higher than in immunocompetent patients. Rates of progression and recurrence following treatment are higher too, in part due to the effects of the immunosu......Background The incidence of actinic keratoses (AK) and non-melanoma skin cancer (NMSC) in organ transplant recipients (OTRs) is significantly higher than in immunocompetent patients. Rates of progression and recurrence following treatment are higher too, in part due to the effects...... investigate induced immunosuppression with PDT in healthy volunteers....

  14. Real-world approach to actinic keratosis management

    DEFF Research Database (Denmark)

    Dirschka, Thomas; Gupta, Girish; Micali, Giuseppe

    2017-01-01

    Actinic keratosis (AK) is a chronic skin disease in which multiple clinical and subclinical lesions co-exist across large areas of sun-exposed skin, resulting in field cancerisation. Lesions require treatment because of their potential to transform into invasive squamous cell carcinoma. This arti......Actinic keratosis (AK) is a chronic skin disease in which multiple clinical and subclinical lesions co-exist across large areas of sun-exposed skin, resulting in field cancerisation. Lesions require treatment because of their potential to transform into invasive squamous cell carcinoma...

  15. The integrin-actin connection, an eternal love affair

    DEFF Research Database (Denmark)

    Brakebusch, Cord; Fässler, Reinhard

    2003-01-01

    Integrin receptors connect the extracellular matrix to the actin cytoskeleton. This interaction can be viewed as a cyclical liaison, which develops again and again at new adhesion sites only to cease at sites of de-adhesion. Recent work has demonstrated that multidomain proteins play crucial roles...... in the integrin-actin connection by providing a high degree of regulation adjusted to the needs of the cell. In this review we present several examples of this paradigm and with special emphasis on the ILK-PINCH-parvin complex, which amply demonstrates how structural and signalling functions are linked together....

  16. DNA repair deficiency in lymphocytes from patients with actinic keratosis

    International Nuclear Information System (INIS)

    Abo-Darub, J.M.; Mackie, R.; Pitts, J.D.

    1978-01-01

    DNA repair activity was measured in peripheral blood lymphocytes from 18 patients with Actinic Keratosis and 18 age-matched control subjects, by comparing the incorporation of 3 H-thymidine into cells after irradiation with ultraviolet light with that into unirradiated cells. The incorporation was followed autoradiographically or by measuring acid insoluble radioactivity in cells labelled in the presence of hydroxyurea. The repair activity in lymphocytes from Actinic keratosis patients was only 47.1% (+-6.5%) of that in cells from the control subjects

  17. Functional compartmentalization of the human superficial masseter muscle.

    Directory of Open Access Journals (Sweden)

    Rodrigo A Guzmán-Venegas

    Full Text Available Some muscles have demonstrated a differential recruitment of their motor units in relation to their location and the nature of the motor task performed; this involves functional compartmentalization. There is little evidence that demonstrates the presence of a compartmentalization of the superficial masseter muscle during biting. The aim of this study was to describe the topographic distribution of the activity of the superficial masseter (SM muscle's motor units using high-density surface electromyography (EMGs at different bite force levels. Twenty healthy natural dentate participants (men: 4; women: 16; age 20±2 years; mass: 60±12 kg, height: 163±7 cm were selected from 316 volunteers and included in this study. Using a gnathodynamometer, bites from 20 to 100% maximum voluntary bite force (MVBF were randomly requested. Using a two-dimensional grid (four columns, six electrodes located on the dominant SM, EMGs in the anterior, middle-anterior, middle-posterior and posterior portions were simultaneously recorded. In bite ranges from 20 to 60% MVBF, the EMG activity was higher in the anterior than in the posterior portion (p-value = 0.001.The center of mass of the EMG activity was displaced towards the posterior part when bite force increased (p-value = 0.001. The topographic distribution of EMGs was more homogeneous at high levels of MVBF (p-value = 0.001. The results of this study show that the superficial masseter is organized into three functional compartments: an anterior, a middle and a posterior compartment. However, this compartmentalization is only seen at low levels of bite force (20-60% MVBF.

  18. [Occupational trauma due to superficial corneal foreign body].

    Science.gov (United States)

    Gerente, Vanessa Miroski; Melo, Gustavo Barreto de; Regatieri, Caio Vinícius Saito; Alvarenga, Lênio Souza; Martins, Elizabeth Nogueira

    2008-01-01

    To evaluate the epidemiology of superficial corneal foreign body. Patients who were seen at the Emergency Service of the Federal University of São Paulo, from April/05 to June/05, were screened and those with superficial corneal foreign body were interviewed. Data regarding gender, age, occupation, employment status, availability and use of protective devices and supervision of their use were collected. Awareness of the possible complications was also assessed. Results were analyzed using chi-square or Fisher exact test. One hundred twenty-three patients were interviewed. Only 3 patients were female. The mean age was 36 years. Most injuries occurred at the workplace (86.2%), and 58.4% of the patients did not have a legal employment registration. The occupational activities most frequently reported were construction related activities (44.3%) and welding/soldering (11.3%). In most workplaces (79.8%) protective devices were available and 85.3% of the patients were instructed to use them. A safety device was being used during the accident in 34.2% of the cases and this was more frequent among patients that had legal employment registry (p=0.008) and among those under supervision (p=0.0415). The majority of the patients (68.9%) were aware of the risk of severe complications. Most patients with superficial corneal foreign body are aware of its severe complications and injuries usually occur in places where safety devices are available and often during their use. Our findings suggest that prevention should focus on supervision and correct use of safety devices.

  19. Superficial thrombophlebitis (Mondor′s Disease after breast augmentation surgery

    Directory of Open Access Journals (Sweden)

    Viana Giovanni Andre

    2008-01-01

    Full Text Available Although the aetiology of Mondor′s disease remains unclear, the most commonly cited cause is trauma of some sort. Although surgical trauma has frequently been quoted, reports that specifically implicate aesthetic breast surgery are unusual in the literature. In this article, the authors report a case of superficial thrombophlebitis of the anterolateral chest wall secondary to breast augmentation surgery in a woman, five months after the procedure. The authors performed an analysis of the disease′s main etiologic components and preponderant clinical aspects, and determined all appropriate therapeutic measures.

  20. High dose rate brachytherapy for superficial cancer of the esophagus

    International Nuclear Information System (INIS)

    Maingon, Philippe; D'Hombres, Anne; Truc, Gilles; Barillot, Isabelle; Michiels, Christophe; Bedenne, Laurent; Horiot, Jean Claude

    2000-01-01

    Purpose: We analyzed our experience with external radiotherapy, combined modality treatment, or HDR brachytherapy alone to limited esophageal cancers. Methods and Materials: From 1991 to 1996, 25 patients with limited superficial esophagus carcinomas were treated by high dose rate brachytherapy. The mean age was 63 years (43-86 years). Five patients showed superficial local recurrence after external radiotherapy. Eleven patients without invasion of the basal membrane were staged as Tis. Fourteen patients with tumors involving the submucosa without spreading to the muscle were staged as T1. Treatment consisted of HDR brachytherapy alone in 13 patients, external radiotherapy and brachytherapy in 8 cases, and concomitant chemo- and radiotherapy in 4 cases. External beam radiation was administered to a total dose of 50 Gy using 2 Gy daily fractions in 5 weeks. In cases of HDR brachytherapy alone (13 patients), 6 applications were performed once a week. Results: The mean follow-up is 31 months (range 24-96 months). Twelve patients received 2 applications and 13 patients received 6 applications. Twelve patients experienced a failure (48%), 11/12 located in the esophagus, all of them in the treated volume. One patient presented an isolated distant metastasis. In the patients treated for superficial recurrence, 4/5 were locally controlled (80%) by brachytherapy alone. After brachytherapy alone, 8/13 patients were controlled (61%). The mean disease-free survival is 14 months (1-36 months). Overall survival is 76% at 1 year, 37% at 2 years, and 14% at 3 years. Overall survival for Tis patients is 24% vs. 20% for T1 (p 0.83). Overall survival for patients treated by HDR brachytherapy alone is 43%. One patient presented with a fistula with local failure after external radiotherapy and brachytherapy. Four stenosis were registered, two were diagnosed on barium swallowing without symptoms, and two required dilatations. Conclusion: High dose rate brachytherapy permits the treating

  1. Modelos de mantenimiento y reemplazo de equipos en mineria superficial

    OpenAIRE

    Llanque Maquera, Oscar Eloy; Llanque Maquera, Oscar Eloy

    1999-01-01

    El mantenimiento y reemplazamiento de equipos en Minería Superficie tiene una gran importada por el mismo hecho que se mueven volumen de material y por las altas inversiones hechas en maquinaria, a su vez por la influencia en los costos de operación. Por tal razón es el interés en desarrollar los modelos de mantenimiento y reemplazamiento de equipos en minería e implemento el mantenimiento predictivo, como una actividad complementaria a los mantenimientos que se realizan comúnmente como son p...

  2. Actin Filaments and Myosin I Alpha Cooperate with Microtubules for the Movement of LysosomesV⃞

    OpenAIRE

    Cordonnier, Marie-Neige; Dauzonne, Daniel; Louvard, Daniel; Coudrier, Evelyne

    2001-01-01

    An earlier report suggested that actin and myosin I alpha (MMIα), a myosin associated with endosomes and lysosomes, were involved in the delivery of internalized molecules to lysosomes. To determine whether actin and MMIα were involved in the movement of lysosomes, we analyzed by time-lapse video microscopy the dynamic of lysosomes in living mouse hepatoma cells (BWTG3 cells), producing green fluorescent protein actin or a nonfunctional domain of MMIα. In GFP-actin cells, lysosomes displayed ...

  3. Fragmentation of Human Erythrocyte Actin following Exposure to Hypoxia

    Czech Academy of Sciences Publication Activity Database

    Risso, A.; Santamaria, B.; Pistarino, E.; Cosulich, M. E.; Pompach, Petr; Bezouška, Karel; Antonutto, G.

    2010-01-01

    Roč. 123, č. 1 (2010), s. 6-13 ISSN 0001-5792 Institutional research plan: CEZ:AV0Z50200510 Keywords : beta-Actin * Erythrocytes * Hypoxia Subject RIV: EE - Microbiology, Virology Impact factor: 1.316, year: 2010

  4. The roles of the actin cytoskeleton in fear memory formation

    Directory of Open Access Journals (Sweden)

    Raphael eLamprecht

    2011-07-01

    Full Text Available The formation and storage of fear memory is needed to adapt behavior and avoid danger during subsequent fearful events. However, fear memory may also play a significant role in stress and anxiety disorders. When fear becomes disproportionate to that necessary to cope with a given stimulus, or begins to occur in inappropriate situations, a fear or anxiety disorder exists. Thus, the study of cellular and molecular mechanisms underpinning fear memory may shed light on the formation of memory and on anxiety and stress related disorders. Evidence indicates that fear learning leads to changes in neuronal synaptic transmission and morphology in brain areas underlying fear memory formation including the amygdala and hippocampus. The actin cytoskeleton has been shown to participate in these key neuronal processes. Recent findings show that the actin cytoskeleton is needed for fear memory formation and extinction. Moreover, the actin cytoskeleton is involved in synaptic plasticity and in neuronal morphogenesis in brain areas that mediate fear memory. The actin cytoskeleton may therefore mediate between synaptic transmission during fear learning and long-term cellular alterations mandatory for fear memory formation.

  5. Force Exertion and Transmission in Cross-Linked Actin Networks

    Science.gov (United States)

    Stam, Samantha

    Cells are responsive to external cues in their environment telling them to proliferate or migrate within their surrounding tissue. Sensing of cues that are mechanical in nature, such stiffness of a tissue or forces transmitted from other cells, is believed to involve the cytoskeleton of a cell. The cytoskeleton is a complex network of proteins consisting of polymers that provide structural support, motor proteins that remodel these structures, and many others. We do not yet have a complete understanding of how cytoskeletal components respond to either internal or external mechanical force and stiffness. Such an understanding should involve mechanisms by which constituent molecules, such as motor proteins, are responsive to mechanics. Additionally, physical models of how forces are transmitted through biopolymer networks are necessary. My research has focused on networks formed by the cytoskeletal filament actin and the molecular motor protein myosin II. Actin filaments form networks and bundles that form a structural framework of the cell, and myosin II slides actin filaments. In this thesis, we show that stiffness of an elastic load that opposes myosin-generated actin sliding has a very sharp effect on the myosin force output in simulations. Secondly, we show that the stiffness and connectivity of cytoskeletal filaments regulates the contractility and anisotropy of network deformations that transmit force on material length scales. Together, these results have implications for predicting and interpreting the deformations and forces in biopolymeric active materials.

  6. Actinic cell effects after radiotherapy for cervical cancer

    International Nuclear Information System (INIS)

    Padilha, C.M.L.; Bergmann, A.; Chaves, C.B.P.; Thuler, L.C.S.; Araújo Junior, M.L.C.; Souza, S.A.L. de

    2017-01-01

    Introduction: It is very common for patients with cervical cancer to be referred to the radiotherapy when the disease is in advanced stages, this fact determines high rates of locoregional recurrence. Radiation treatment causes actinic morphological changes, not only in neoplastic epithelial cells, but also in normal cells. These changes induced by radiation, often make difficult the differential diagnosis of the residual lesion, resulting in a dilemma in cytopathological follow-up. Objective: To describe the actinic cytopathologic changes in patients submitted to radiotherapy for cervical cancer. Methodology: The re-evaluation of cytopathologic smears and description of actinic cytopathic effects were performed. This information was complemented by the cytopathological report of the smears, available in the archives of the Division of Pathology (DIPAT) / INCA. Results: The most frequent cytopathological changes observed were: nuclear activation, cytoplasmic enhancement, cytoplasmic vacuolisation, eosinophilia, polychromasia, multinucleated giant cells, binucleation, nuclear vacuolisation, prominent nucleoli, as well as presence of leukocyte exudate. Conclusion: The cytopathological diagnosis of tumor persistence or recurrence after radiotherapy is always a great challenge for the professional, even the experienced one. Studies and reports in the literature on actinic cytopathologic changes and radiotherapy are scarce

  7. Actinic Keratosis and Non-Invasive Diagnostic Techniques: An Update

    Science.gov (United States)

    Casari, Alice; Chester, Johanna; Pellacani, Giovanni

    2018-01-01

    Actinic keratosis represents the earliest manifestation of non-melanoma skin cancer. Because of their risk of progression to invasive squamous cell carcinoma, an earlier diagnosis and treatment are mandatory. Their diagnosis sometimes could represent a challenge even for expert dermatologists. Dermoscopy, confocal laser microscopy and optical coherence tomography could help clinicians in diagnosis. PMID:29316678

  8. Pharmacoeconomic Considerations in Treating Actinic Keratosis: An Update.

    Science.gov (United States)

    Vale, Spencer M; Hill, Dane; Feldman, Steven R

    2017-02-01

    Actinic keratosis is one of the most common dermatological diagnoses worldwide, especially among the elderly, fair-skinned, and immunocompromised, and is associated with a risk of transformation to skin cancer. With actinic keratosis and skin cancer prevalence increasing as the aged population expands in the US, optimizing treatment strategies may produce cost savings for the healthcare system. Since the time of our last review in 2008, investigation of the economic considerations in treating actinic keratosis has advanced. To provide an update of treatment cost effectiveness and to review factors relating to the costs of care, we conducted a systematic review of pharmacoeconomic publications since December 2008. We identified 11 pharmacoeconomic studies, with one cost-of-treatment, five cost-effectiveness, and five cost-utility analyses. Photodynamic therapy (PDT) was well tolerated and produced a favorable cosmetic outcome in most studies. Ingenol mebutate, the newest but most expensive topical field therapy, 5-fluorouracil, and PDT were the most cost-effective treatments in our review. Patient adherence to therapy and the management of adverse effects were significant contributors to treatment costs. In the US, treatment guidelines and formalized cost-effectiveness analyses for actinic keratosis are absent from the recent literature. Future pharmacoeconomic investigation will depend on up-to-date comparative efficacy data, as well as clarification of rates of, and management strategies for, adverse effects, therapeutic non-adherence, and lesion recurrence.

  9. Decidable and undecidable arithmetic functions in actin filament networks

    Science.gov (United States)

    Schumann, Andrew

    2018-01-01

    The plasmodium of Physarum polycephalum is very sensitive to its environment, and reacts to stimuli with appropriate motions. Both the sensory and motor stages of these reactions are explained by hydrodynamic processes, based on fluid dynamics, with the participation of actin filament networks. This paper is devoted to actin filament networks as a computational medium. The point is that actin filaments, with contributions from many other proteins like myosin, are sensitive to extracellular stimuli (attractants as well as repellents), and appear and disappear at different places in the cell to change aspects of the cell structure—e.g. its shape. By assembling and disassembling actin filaments, some unicellular organisms, like Amoeba proteus, can move in response to various stimuli. As a result, these organisms can be considered a simple reversible logic gate—extracellular signals being its inputs and motions its outputs. In this way, we can implement various logic gates on amoeboid behaviours. These networks can embody arithmetic functions within p-adic valued logic. Furthermore, within these networks we can define the so-called diagonalization for deducing undecidable arithmetic functions.

  10. Actin and myosin contribute to mammalian mitochondrial DNA maintenance

    Science.gov (United States)

    Reyes, A.; He, J.; Mao, C. C.; Bailey, L. J.; Di Re, M.; Sembongi, H.; Kazak, L.; Dzionek, K.; Holmes, J. B.; Cluett, T. J.; Harbour, M. E.; Fearnley, I. M.; Crouch, R. J.; Conti, M. A.; Adelstein, R. S.; Walker, J. E.; Holt, I. J.

    2011-01-01

    Mitochondrial DNA maintenance and segregation are dependent on the actin cytoskeleton in budding yeast. We found two cytoskeletal proteins among six proteins tightly associated with rat liver mitochondrial DNA: non-muscle myosin heavy chain IIA and β-actin. In human cells, transient gene silencing of MYH9 (encoding non-muscle myosin heavy chain IIA), or the closely related MYH10 gene (encoding non-muscle myosin heavy chain IIB), altered the topology and increased the copy number of mitochondrial DNA; and the latter effect was enhanced when both genes were targeted simultaneously. In contrast, genetic ablation of non-muscle myosin IIB was associated with a 60% decrease in mitochondrial DNA copy number in mouse embryonic fibroblasts, compared to control cells. Gene silencing of β-actin also affected mitochondrial DNA copy number and organization. Protease-protection experiments and iodixanol gradient analysis suggest some β-actin and non-muscle myosin heavy chain IIA reside within human mitochondria and confirm that they are associated with mitochondrial DNA. Collectively, these results strongly implicate the actomyosin cytoskeleton in mammalian mitochondrial DNA maintenance. PMID:21398640

  11. Actin-Dependent Alterations of Dendritic Spine Morphology in Shankopathies

    Directory of Open Access Journals (Sweden)

    Tasnuva Sarowar

    2016-01-01

    Full Text Available Shank proteins (Shank1, Shank2, and Shank3 act as scaffolding molecules in the postsynaptic density of many excitatory neurons. Mutations in SHANK genes, in particular SHANK2 and SHANK3, lead to autism spectrum disorders (ASD in both human and mouse models. Shank3 proteins are made of several domains—the Shank/ProSAP N-terminal (SPN domain, ankyrin repeats, SH3 domain, PDZ domain, a proline-rich region, and the sterile alpha motif (SAM domain. Via various binding partners of these domains, Shank3 is able to bind and interact with a wide range of proteins including modulators of small GTPases such as RICH2, a RhoGAP protein, and βPIX, a RhoGEF protein for Rac1 and Cdc42, actin binding proteins and actin modulators. Dysregulation of all isoforms of Shank proteins, but especially Shank3, leads to alterations in spine morphogenesis, shape, and activity of the synapse via altering actin dynamics. Therefore, here, we highlight the role of Shank proteins as modulators of small GTPases and, ultimately, actin dynamics, as found in multiple in vitro and in vivo models. The failure to mediate this regulatory role might present a shared mechanism in the pathophysiology of autism-associated mutations, which leads to dysregulation of spine morphogenesis and synaptic signaling.

  12. Interconnection between actin cytoskeleton and plant defense signaling

    Czech Academy of Sciences Publication Activity Database

    Janda, Martin; Matoušková, J.; Burketová, Lenka; Valentová, O.

    2014-01-01

    Roč. 9, č. 11 (2014) ISSN 1559-2316 R&D Projects: GA ČR(CZ) GAP501/11/1654 Institutional support: RVO:61389030 Keywords : Actin * Cytoskeleton * Pathogen Subject RIV: ED - Physiology http://gateway.isiknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=Alerting&SrcApp=Alerting&DestApp=MEDLINE&DestLinkType=FullRecord&UT=25482795

  13. Suspected Pulmonary Metastasis of Actinic Cutaneous Squamous Cell Carcinoma

    Directory of Open Access Journals (Sweden)

    Monet E. Meter

    2017-01-01

    Full Text Available Introduction. It is rare for actinic or squamous cell carcinoma (SCC in situ to metastasize. Case Presentation. A 67-year-old male had a significant medical history including severe psoriatic arthritis treated with UVB, methotrexate, and rapamycin. He had twenty-five different skin excisions of actinic keratosis four of which were invasive SCC. Our patient developed shortness of breath necessitating a visit to the emergency department. A CT scan of his chest revealed a mass in the right lower lung. A subsequent biopsy of the mass revealed well-differentiated SCC. He underwent thoracoscopic surgery with wedge resection of the lung lesion. Discussion. Actinic keratosis (AK is considered precancerous and associated with UV exposure. It exists as a continuum of progression with low potential for malignancy. The majority of invasive SCCs are associated with malignant progression of AK, but only 5–10% of AKs will progress to malignant potential. Conclusion. In this case, a new finding of lung SCC in the setting of multiple invasive actinic cutaneous SCC associated with a history of extensive UV light exposure and immunosuppression supports a metastatic explanation for lung cancer.

  14. Genomic instability in human actinic keratosis and squamous cell carcinoma

    Science.gov (United States)

    Cabral, Luciana Sanches; Neto, Cyro Festa; Sanches, José A; Ruiz, Itamar R G

    2011-01-01

    OBJECTIVE: To compare the repetitive DNA patterns of human actinic keratoses and squamous cell carcinomas to determine the genetic alterations that are associated with malignant transformation. INTRODUCTION: Cancer cells are prone to genomic instability, which is often due to DNA polymerase slippage during the replication of repetitive DNA and to mutations in the DNA repair genes. The progression of benign actinic keratoses to malignant squamous cell carcinomas has been proposed by several authors. MATERIAL AND METHODS: Eight actinic keratoses and 24 squamous cell carcinomas (SCC), which were pair-matched to adjacent skin tissues and/or leucocytes, were studied. The presence of microsatellite instability (MSI) and the loss of heterozygosity (LOH) in chromosomes 6 and 9 were investigated using nine PCR primer pairs. Random Amplified Polymorphic DNA patterns were also evaluated using eight primers. RESULTS: MSI was detected in two (D6S251, D9S50) of the eight actinic keratosis patients. Among the 8 patients who had squamous cell carcinoma-I and provided informative results, a single patient exhibited two LOH (D6S251, D9S287) and two instances of MSI (D9S180, D9S280). Two LOH and one example of MSI (D6S251) were detected in three out of the 10 patients with squamous cell carcinoma-II. Among the four patients with squamous cell carcinoma-III, one patient displayed three MSIs (D6S251, D6S252, and D9S180) and another patient exhibited an MSI (D9S280). The altered random amplified polymorphic DNA ranged from 70% actinic keratoses, 76% squamous cell carcinoma-I, and 90% squamous cell carcinoma-II, to 100% squamous cell carcinoma-III. DISCUSSION: The increased levels of alterations in the microsatellites, particularly in D6S251, and the random amplified polymorphic DNA fingerprints were statistically significant in squamous cell carcinomas, compared with actinic keratoses. CONCLUSION: The overall alterations that were observed in the repetitive DNA of actinic keratoses and

  15. Probing cytoplasmic organization and the actin cytoskeleton of plant cells with optical tweezers

    NARCIS (Netherlands)

    Ketelaar, T.; Honing, van der H.S.; Emons, A.M.C.

    2010-01-01

    In interphase plant cells, the actin cytoskeleton is essential for intracellular transport and organization. To fully understand how the actin cytoskeleton functions as the structural basis for cytoplasmic organization, both molecular and physical aspects of the actin organization have to be

  16. Actin is closely associated with RNA polymerase II and involved in activation of gene transcription

    International Nuclear Information System (INIS)

    Zhu Xiaojuan; Zeng Xianlu; Huang Baiqu; Hao, Shui

    2004-01-01

    Biochemical and morphological studies have demonstrated the presence of actin in the nucleus of different eukaryotic cells, whereas its role remains unclear. In this work, we studied the interaction and the functional relationship between nuclear actin and RNA polymerase II (RNAP II). The immunofluorescence study demonstrated a clear co-localization of nuclear actin with RNAP II in HeLa cells. Meanwhile, actin can be immunoprecipitated by anti-RNAP II antibody, indicating that they could interact with each other. Treatment of cells with α-amanitin induced the formation of actin bundle network in the nucleoplasm. Blocking of the formation of filamentous actin (F-actin) by cytochalasin B modified the distribution of actin. Although the actin content remained unchanged in resting and concanavalinA stimulated mouse lymphocytes, the actin content in the nuclei showed a progressive increase after stimulation. Furthermore, the antibody against actin blocked RNA synthesis in a eukaryotic in vitro transcription system. These observations implicate that nuclear actin interacts with RNAP II and may have function on the RNAP II-mediated transcription

  17. Multidrug Resistance-Related Protein 1 (MRP1) Function and Localization Depend on Cortical Actin

    NARCIS (Netherlands)

    Hummel, Ina; Klappe, Karin; Ercan, Cigdem; Kok, Jan Willem

    MRP1 (ABCC1) is known to be localized in lipid rafts. Here we show in two different cell lines that localization of Mrp1/MRP1 (Abcc1/ABCC1) in lipid rafts and its function as an efflux pump are dependent on cortical actin. Latrunculin B disrupts both cortical actin and actin stress fibers. This

  18. The effect of pyrene labelling on the thermal stability of actin filaments

    International Nuclear Information System (INIS)

    Halasi, Szulamit; Papp, Gabor; Bugyi, Beata; Barko, Szilvia; Orban, Jozsef; Ujfalusi, Zoltan; Visegrady, Balazs

    2006-01-01

    The ability of actin to form filaments is fundamental to its biological function and often characterised by various methods in vitro. One of the most frequently used methods capitalises on the observation that the fluorescence emission of a pyrene label on the Cys-374 residue of actin is enhanced by a factor of ∼20 during polymerisation. This method inherently involves the chemical modification of actin monomers with pyrene. It was reported earlier that the pyrene labelling of actin monomers has only small effect on the polymerisation and depolymerisation rates of actin, indicating that the method is suitable to characterise the effect of actin-binding proteins or peptides on the polymerisation kinetics. In our present work we tested the effect of the pyrene labelling on the thermal denaturation of actin filaments by using the method of differential scanning calorimetry (DSC). By recording the heat denaturation profiles of unlabelled and pyrene labelled actin filaments we observed that pyrene labelling shifted the melting point (T m ) of actin filaments from 66 to 68 deg. C. A similar effect was detected in the presence of equimolar concentration of phalloidin where the T m shifted from 79 to 82 deg. C. We concluded that the observed pyrene labelling induced differences of the thermal denaturation of actin filaments were small. The DSC results, therefore, confirmed that the methods based on the measurements of pyrene intensity during actin polymerisation are suitable to characterise the polymerisation kinetics of actin under in vitro conditions

  19. Optical coherence tomography and polarimetry of superficial skin biopsies

    Directory of Open Access Journals (Sweden)

    S. R. Utz

    2015-01-01

    Full Text Available The goal. Of this study was to develop and assess the efficacy of polarization probing of biotissues in vitro. The method is based on the determination of polarization parameters of scattered radiation. Materials and methods. The well-known superficial epidermis stripping method was applied using the Sulfacrylate self-sterile medical adhesive. Small portions of thin layers of the adhesive were applied to slide plates and then to different skin sites. The corneous layer in the normal condition and in case of skin diseases (psoriasis, lichen acuminatus, discoid lupus erythematosus, alopecia, itching and demodectic mange was examined based on the optical coherence tomography (OCT method using the 0CS1300SS device (Thorlabs Inc, USA. Results. The authors obtained pictures visualizing the structural organization of different layers of the epidermis using the superficial epidermis biopsy method in case of lichen acuminatus, hyperkeratosis, itching and other skin diseases. Conclusion. This method ensures non-invasive high-precision measurement of the structure of different layers of the epidermis, which may be useful both for research purposes and practical dermatology.

  20. Superficially located enlarged lymphoid follicles characterise nodular gastritis.

    Science.gov (United States)

    Okamura, Takuma; Sakai, Yasuhiro; Hoshino, Hitomi; Iwaya, Yugo; Tanaka, Eiji; Kobayashi, Motohiro

    2015-01-01

    Nodular gastritis is a form of chronic Helicobacter pylori gastritis affecting the gastric antrum and characterised endoscopically by the presence of small nodular lesions resembling gooseflesh. It is generally accepted that hyperplasia of lymphoid follicles histologically characterises nodular gastritis; however, quantitative analysis in support of this hypothesis has not been reported. Our goal was to determine whether nodular gastritis is characterised by lymphoid follicle hyperplasia.The number, size, and location of lymphoid follicles in nodular gastritis were determined and those properties compared to samples of atrophic gastritis. The percentages of high endothelial venule (HEV)-like vessels were also evaluated.The number of lymphoid follicles was comparable between nodular and atrophic gastritis; however, follicle size in nodular gastritis was significantly greater than that seen in atrophic gastritis. Moreover, lymphoid follicles in nodular gastritis were positioned more superficially than were those in atrophic gastritis. The percentage of MECA-79 HEV-like vessels was greater in areas with gooseflesh-like lesions in nodular versus atrophic gastritis.Superficially located hyperplastic lymphoid follicles characterise nodular gastritis, and these follicles correspond to gooseflesh-like nodular lesions observed endoscopically. These observations suggest that MECA-79 HEV-like vessels could play at least a partial role in the pathogenesis of nodular gastritis.

  1. Lactic acid peeling in superficial acne scarring in Indian skin.

    Science.gov (United States)

    Sachdeva, Silonie

    2010-09-01

    Chemical peeling with both alpha and beta hydroxy acids has been used to improve acne scarring with pigmentation. Lactic acid, a mild alpha hydroxy acid, has been used in the treatment of various dermatological indications but no study is reported in acne scarring with pigmentation. To evaluate the efficacy and safety of full strength pure lactic acid 92% (pH 2.0) chemical peel in superficial acne scarring in Indian skin. Seven patients, Fitzpatrick skin type IV-V, in age group 20-30 years with superficial acne scarring were enrolled in the study. Chemical peeling was done with lactic acid at an interval of 2 weeks to a maximum of four peels. Pre- and post-peel clinical photographs were taken at every session. Patients were followed every month for 3 months after the last peel to evaluate the effects. At the end of 3 months, there was definite improvement in the texture, pigmentation, and appearance of the treated skin, with lightening of scars. Significant improvement (greater than 75% clearance of lesions) occurred in one patient (14.28%), good improvement (51-75% clearance) in three patients (42.84%), moderate improvement (26-50% clearance) in two patients (28.57%), and mild improvement (1-25% clearance) in one patient (14.28%). © 2010 Wiley Periodicals, Inc.

  2. Superficial corneal crosslinking during laser in situ keratomileusis.

    Science.gov (United States)

    Seiler, Theo G; Fischinger, Isaak; Koller, Tobias; Derhartunian, Viktor; Seiler, Theo

    2015-10-01

    To determine the safety of superficial corneal crosslinking after laser in situ keratomileusis (LASIK). Institut für Refraktive und Ophthalmo-Chirurgie, Zurich, Switzerland. Prospective study. Eyes with an ectasia risk score of 2 or higher were treated with standard LASIK (90 μm flap) for myopia correction, after which a rapid corneal crosslinking was performed in the interface (riboflavin 0.5% for 2 minutes, 9 mW/cm(2) for 5 minutes) (Group 1). The follow-up was up to 1 year. The prevalence of complications was statistically compared with that in a group of eyes matched regarding age, sex, and attempted refractive correction that were treated with standard LASIK only (Group 2). One month postoperatively, 5 eyes in Group 1 lost 1 line of corrected distance visual acuity (CDVA) compared with 1 eye in Group 2 (P rate of less than 5%. The refractive success was identical in both groups. Early postoperative complications such as erosions (16%), diffuse lamellar keratitis (DLK) stage 1 (38%), and DLK stage 2 (5%) were statistically significantly more frequent after superficial corneal crosslinking, leading to a statistically significantly reduced uncorrected distance visual acuity at 1 month (P interest in any material or method mentioned. Copyright © 2015 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

  3. Local superficial hyperthermia in combination with low-dose radiation therapy for palliation of superficially localized metastases

    International Nuclear Information System (INIS)

    Owczarek, G.; Miszczyk, L.

    2005-01-01

    Full text: The aim of this study is to evaluate the response of superficially located metastases and local toxicity to microwave hyperthermia combined with radiation therapy. From May 2003 through December 2004 58 patients (33 male, 25 female; mean age 60 years) with lymph nodes or skin metastases were treated with microwave superficial hyperthermia combined with low-dose radiation therapy. Hyperthermia was administered twice weekly with high frequency applicator (∼900 Mhz) with water bolus. The temperature was set to 43 o C for 45 minutes. Radiotherapy was performed daily with dose 2 Gy or 4 Gy per fraction, to a total dose 20 Gy. There were 47 patients with carcinoma, 4 with sarcoma, 7 with melanoma. Treated regions were: head and neck (37 patients), chest wall 8, abdomen wall and groins 4, upper and lower limb 2 and 8 patients respectively. Primary tumor sites were: head and neck region (9 patients), lung 15, alimentary tract 8, breast 5, soft tissue 8, urogenital 4 and 9 patients with primary tumor site unknown. The toxicity was evaluated using 6 step scale: 0-no skin reaction, 1-faint red mark, 2-distinct red mark, 3-blisters, 4-brown mark, 5-necrosis. Presence of pain and its intensity were also analyzed. Diameter of tumor after the treatment was observed. Complete response was achieved in 5 patients (8.5 %), and partial response in 29 patients (50 %), no response was observed in 12 patients (20 %) and progression of tumor in 7 patients (12 %). No skin reaction was observed in 3 patients, faint red mark in 14 patients, distinct red mark in 28 patients, blisters in 8 patients, brown mark in 4 patients and necrosis in 1 patient. The pain occurred in 9 patients but it was no the cause of stopping treatment. Local superficial hyperthermia combined with low-dose radiation therapy is an effective method of treatment in a proportion of patients with superficial metastases. This combination of treatment modalities is well tolerated and is useful for palliation

  4. Actin grips: circular actin-rich cytoskeletal structures that mediate the wrapping of polymeric microfibers by endothelial cells.

    Science.gov (United States)

    Jones, Desiree; Park, DoYoung; Anghelina, Mirela; Pécot, Thierry; Machiraju, Raghu; Xue, Ruipeng; Lannutti, John J; Thomas, Jessica; Cole, Sara L; Moldovan, Leni; Moldovan, Nicanor I

    2015-06-01

    Interaction of endothelial-lineage cells with three-dimensional substrates was much less studied than that with flat culture surfaces. We investigated the in vitro attachment of both mature endothelial cells (ECs) and of less differentiated EC colony-forming cells to poly-ε-capro-lactone (PCL) fibers with diameters in 5-20 μm range ('scaffold microfibers', SMFs). We found that notwithstanding the poor intrinsic adhesiveness to PCL, both cell types completely wrapped the SMFs after long-term cultivation, thus attaining a cylindrical morphology. In this system, both EC types grew vigorously for more than a week and became increasingly more differentiated, as shown by multiplexed gene expression. Three-dimensional reconstructions from multiphoton confocal microscopy images using custom software showed that the filamentous (F) actin bundles took a conspicuous ring-like organization around the SMFs. Unlike the classical F-actin-containing stress fibers, these rings were not associated with either focal adhesions or intermediate filaments. We also demonstrated that plasma membrane boundaries adjacent to these circular cytoskeletal structures were tightly yet dynamically apposed to the SMFs, for which reason we suggest to call them 'actin grips'. In conclusion, we describe a particular form of F-actin assembly with relevance for cytoskeletal organization in response to biomaterials, for endothelial-specific cell behavior in vitro and in vivo, and for tissue engineering. Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. The superficial ulnar artery: development and clinical significance Artéria ulnar superficial: desenvolvimento e relevância clínica

    Directory of Open Access Journals (Sweden)

    Srinivasulu Reddy

    2007-09-01

    Full Text Available The principal arteries of the upper limb show a wide range of variation that is of considerable interest to orthopedic surgeons, plastic surgeons, radiologists and anatomists. We present here a case of superficial ulnar artery found during the routine dissection of right upper limb of a 50-year-old male cadaver. The superficial ulnar artery originated from the brachial artery, crossed the median nerve anteriorly and ran lateral to this nerve and the brachial artery. The superficial ulnar artery in the arm gave rise to a narrow muscular branch to the biceps brachii. At the elbow level the artery ran superficial to the bicipital aponeurosis where it was crossed by the median cubital vein. It then ran downward and medially superficial to the forearm flexor muscles, and then downward to enter the hand. At the palm, it formed the superficial and deep palmar arches together with the branches of the radial artery. The presence of a superficial ulnar artery is clinically important when raising forearm flaps in reconstructive surgery. The embryology and clinical significance of the variation are discussed.As principais artérias do membro superior apresentam uma ampla variação, que é relativamente importante a cirurgiões ortopédicos e plásticos, radiologistas e anatomistas.Apresentamosumcaso de artéria ulnar superficial encontrada durante dissecção de rotina de membro superior direito de um cadáver masculino de 50 anos de idade.Aartéria ulnar superficial originava-se da artéria braquial, cruzava o nervo mediano anteriormente e percorria lateralmente esse nervo e a artéria braquial. A artéria ulnar superficial no braço deu origem a um ramo muscular estreito do músculo bíceps braquial. Ao nível do cotovelo, a artéria percorria superficialmente a aponeurose bicipital, onde era cruzada pela veia cubital mediana. Percorria, então, em sentido descendente e medialmente superficial aos músculos flexores do antebraço, e então descendia para

  6. Recruitment Kinetics of Tropomyosin Tpm3.1 to Actin Filament Bundles in the Cytoskeleton Is Independent of Actin Filament Kinetics.

    Science.gov (United States)

    Appaduray, Mark A; Masedunskas, Andrius; Bryce, Nicole S; Lucas, Christine A; Warren, Sean C; Timpson, Paul; Stear, Jeffrey H; Gunning, Peter W; Hardeman, Edna C

    2016-01-01

    The actin cytoskeleton is a dynamic network of filaments that is involved in virtually every cellular process. Most actin filaments in metazoa exist as a co-polymer of actin and tropomyosin (Tpm) and the function of an actin filament is primarily defined by the specific Tpm isoform associated with it. However, there is little information on the interdependence of these co-polymers during filament assembly and disassembly. We addressed this by investigating the recovery kinetics of fluorescently tagged isoform Tpm3.1 into actin filament bundles using FRAP analysis in cell culture and in vivo in rats using intracellular intravital microscopy, in the presence or absence of the actin-targeting drug jasplakinolide. The mobile fraction of Tpm3.1 is between 50% and 70% depending on whether the tag is at the C- or N-terminus and whether the analysis is in vivo or in cultured cells. We find that the continuous dynamic exchange of Tpm3.1 is not significantly impacted by jasplakinolide, unlike tagged actin. We conclude that tagged Tpm3.1 may be able to undergo exchange in actin filament bundles largely independent of the assembly and turnover of actin.

  7. F-actin distribution and function during sexual differentiation in Schizosaccharomyces pombe

    DEFF Research Database (Denmark)

    Petersen, J; Nielsen, O; Egel, R

    1998-01-01

    Sexual differentiation in Schizosaccharomyces pombe is induced from the G1 phase of the cell cycle by nitrogen starvation and the presence of mating pheromones. We describe the distribution of F-actin during sexual differentiation. Cortical F-actin dots have previously been shown to be restricted...... to one end of the rod shaped cell during the G1 phase of the cell cycle. Within half an hour of nitrogen starvation the distribution of cortical F-actin dots switched from being monopolar to bipolar. This was then reversed as the F-actin cytoskeleton repolarized so that cortical F-actin dots accumulated...

  8. Review of photodynamic therapy with 5-methyl aminolevulinate in actinic keratosis, epidermoid carcinoma and basal cell carcinoma

    International Nuclear Information System (INIS)

    Fallas Moya, Said

    2013-01-01

    A bibliographic review was conduced on the use of 5-methyl aminolevulinate in dermatology, specifically in the treatment of actinic keratosis, epidermoid carcinoma and basal cell carcinoma. The basic fundamentals of photodynamic therapy are described. The preparation and method of use of photodynamic therapy with 5-methyl aminolevulinate (MAL-PDT) are detailed. The clinical studies that were realized with photodynamic therapy for the treatment of actinic keratosis, epidermoid carcinoma and basal cell carcinoma are mentioned. Different photo-inducible agents and other current therapeutic options of first-line are compared. The MAL-PDT has have the advantage of to present less side effects and the same have been more tolerable than liquid nitrogen and 5 fluorouracil. The MAL-PDT has been considered as an effective option for the treatment of Bowen's disease. Invasive epidermoid carcinoma has existed without evidence to support the routine use of this therapeutic. For superficial basal cell carcinoma, the MAL-PDT has presented a high cure rate and transient and manageable side effects in extensive and multiple lesions. The MAL-PDT has been an effective and safe treatment in patients with basal cell carcinoma, for those with less depth of 2mm. The MAL-PDT could play an important role in the field of prevention with immunosuppressed patients, particularly, those that have required transplant and its immunosuppression has been pharmacological. The use or not of the MAL-PDT, should be evaluated individually for each patient and to have suitable characteristics for each disease that was cited in this review. The photodynamic therapy with 5-methyl aminolevulinate has been a therapeutic modality of considerable economy, however, it should be evaluated in the context of number of inquiries and side effects that have offered other therapeutic modalities [es

  9. What factors control the superficial lava dome explosivity?

    Science.gov (United States)

    Boudon, Georges; Balcone-Boissard, Hélène; Villemant, Benoit; Morgan, Daniel J.

    2015-04-01

    Dome-forming eruption is a frequent eruptive style; lava domes result from intermittent, slow extrusion of viscous lava. Most dome-forming eruptions produce highly microcrystallized and highly- to almost totally-degassed magmas which have a low explosive potential. During lava dome growth, recurrent collapses of unstable parts are the main destructive process of the lava dome, generating concentrated pyroclastic density currents (C-PDC) channelized in valleys. These C-PDC have a high, but localized, damage potential that largely depends on the collapsed volume. Sometimes, a dilute ash cloud surge develops at the top of the concentrated flow with an increased destructive effect because it may overflow ridges and affect larger areas. In some cases, large lava dome collapses can induce a depressurization of the magma within the conduit, leading to vulcanian explosions. By contrast, violent, laterally directed, explosions may occur at the base of a growing lava dome: this activity generates dilute and turbulent, highly-destructive, pyroclastic density currents (D-PDC), with a high velocity and propagation poorly dependent on the topography. Numerous studies on lava dome behaviors exist, but the triggering of lava dome explosions is poorly understood. Here, seven dome-forming eruptions are investigated: in the Lesser Antilles arc: Montagne Pelée, Martinique (1902-1905, 1929-1932 and 650 y. BP eruptions), Soufrière Hills, Montserrat; in Guatemala, Santiaguito (1929 eruption); in La Chaîne des Puys, France (Puy de Dome and Puy Chopine eruptions). We propose a new model of superficial lava-dome explosivity based upon a textural and geochemical study (vesicularity, microcrystallinity, cristobalite distribution, residual water contents, crystal transit times) of clasts produced by these key eruptions. Superficial explosion of a growing lava dome may be promoted through porosity reduction caused by both vesicle flattening due to gas escape and syn-eruptive cristobalite

  10. Two Crosslinking Technologies for Superficial Reticular Dermis Injection: A Comparative Ultrasound and Histologic Study

    OpenAIRE

    Micheels, Patrick; Besse, Stéphanie; Sarazin, Didier

    2017-01-01

    Background: Few hyaluronic acid fillers have been developed for superficial injection. Objective: To compare the diffusion and integration properties of cohesive polydensified matrix and Vycross® technology hyaluronic acid fillers with lidocaine following injection into the superficial reticular dermis. Methods and materials: Two subjects received two injections each of cohesive polydensified matrix and Vycross® hyaluronic acid (0.2mL/site) in the superficial reticular dermis of the buttock u...

  11. Myxoid stroma and delicate vasculature of a superficial angiomyxoma give rise to the red planet sign

    OpenAIRE

    Green, Margaret; Logemann, Nichola; Sulit, Daryl J

    2014-01-01

    Superficial angiomyxomas are uncommon benign mesenchymal tumors. They often recur locally if partially removed. This case report demonstrates not only the characteristic pathological findings of a superficial angiomyxoma in a 33- year-old man, but also shows a unique dermatoscopic image, which in our estimation resembles a celestial red planet such as the blood moon seen during a lunar eclipse. We propose to call this the “red planet” sign for a superficial angiomyxoma on dermoscopic examinat...

  12. Myxoid stroma and delicate vasculature of a superficial angiomyxoma give rise to the red planet sign.

    Science.gov (United States)

    Green, Margaret; Logemann, Nichola; Sulit, Daryl J

    2014-09-16

    Superficial angiomyxomas are uncommon benign mesenchymal tumors. They often recur locally if partially removed. This case report demonstrates not only the characteristic pathological findings of a superficial angiomyxoma in a 33- year-old man, but also shows a unique dermatoscopic image, which in our estimation resembles a celestial red planet such as the blood moon seen during a lunar eclipse. We propose to call this the "red planet" sign for a superficial angiomyxoma on dermoscopic examination.

  13. The conserved Tarp actin binding domain is important for chlamydial invasion.

    Directory of Open Access Journals (Sweden)

    Travis J Jewett

    2010-07-01

    Full Text Available The translocated actin recruiting phosphoprotein (Tarp is conserved among all pathogenic chlamydial species. Previous reports identified single C. trachomatis Tarp actin binding and proline rich domains required for Tarp mediated actin nucleation. A peptide antiserum specific for the Tarp actin binding domain was generated and inhibited actin polymerization in vitro and C. trachomatis entry in vivo, indicating an essential role for Tarp in chlamydial pathogenesis. Sequence analysis of Tarp orthologs from additional chlamydial species and C. trachomatis serovars indicated multiple putative actin binding sites. In order to determine whether the identified actin binding domains are functionally conserved, GST-Tarp fusions from multiple chlamydial species were examined for their ability to bind and nucleate actin. Chlamydial Tarps harbored variable numbers of actin binding sites and promoted actin nucleation as determined by in vitro polymerization assays. Our findings indicate that Tarp mediated actin binding and nucleation is a conserved feature among diverse chlamydial species and this function plays a critical role in bacterial invasion of host cells.

  14. Dynamics of actin cables in polarized growth of the filamentous fungus Aspergillus nidulans

    Directory of Open Access Journals (Sweden)

    Anna eBergs

    2016-05-01

    Full Text Available Highly polarized growth of filamentous fungi requires a continuous supply of proteins and lipids to the hyphal tip. This transport is managed by vesicle trafficking via the actin and microtubule cytoskeletons and their associated motor proteins. Particularly, actin cables originating from the hyphal tip are essential for hyphal growth. Although specific marker proteins to visualize actin cables have been developed in filamentous fungi, the exact organization and dynamics of actin cables has remained elusive. Here we visualized actin cables using tropomyosin (TpmA and Lifeact fused to fluorescent proteins in Aspergillus nidulans and studied the dynamics and regulation. GFP tagged TpmA visualized dynamic actin cables formed from the hyphal tip with cycles of elongation and shrinkage. The elongation and shrinkage rates of actin cables were similar and approximately 0.6 μm/s. Comparison of actin markers revealed that high concentrations of Lifeact reduced actin dynamics. Simultaneous visualization of actin cables and microtubules suggests temporally and spatially coordinated polymerization and depolymerization between the two cytoskeletons. Our results provide new insights into the molecular mechanism of ordered polarized growth regulated by actin cables and microtubules.

  15. Understanding the three-dimensional anatomy of the superficial lymphatics of the limbs.

    Science.gov (United States)

    Tourani, Saam S; Taylor, G Ian; Ashton, Mark W

    2014-11-01

    There are minimal data in the current literature regarding the depth of the superficial lymphatic collectors of the limbs in relation to the various subcutaneous tissue layers. Injection, microdissection, radiographic, and histologic studies of the superficial lymphatics and the subcutaneous tissues of 32 limbs from 15 human cadavers were performed. Five layers were consistently identified in the integument of all the upper and lower limb specimens: (1) skin, (2) subcutaneous fat, (3) superficial fascia, (4) loose areolar tissue, and (5) deep fascia. Layer 2 was further divided into superficial (2a) and deep (2c) compartments by a thin, transparent, horizontal septum (layer 2b). The main superficial veins and the superficial nerves coursed in layer 4. The lymphatic collectors were found at layer 2c and layer 4. The use of consistent nomenclature to describe the subcutaneous tissue layers facilitates a greater understanding and discussion of the anatomy. In lymphovenous anastomosis for the treatment of lymphedema, indocyanine green lymphography is an unreliable method for identification of the superficial collectors of the thigh. The medial proximal leg, the dorsum of the wrist over the anatomical snuffbox, and the volar proximal forearm provide suitable areas for locating superficial collectors with nearby matching size veins. In vertical medial thigh lift, choosing a dissection plane superficial to the great saphenous vein is unlikely to preserve the collectors of the ventromedial bundle.

  16. Superficies de titanio en implantes dentales de carga inmediata

    OpenAIRE

    Aparicio, C.; Padrós, A.; Planell Estany, Josep Anton; Gil, F.J.

    2008-01-01

    Se ha obtenido una capa de fosfato de calcio del mismo contenido mineral que el hueso en la superficie del implante dental de titanio, que tiene como consecuencia una muy rápida y eficaz osteointegración. Esta capa se ha obtenido mediante un método biomimético por vía termoquímica. La capa se une al substrato por enlace químico. Este hecho evita todos los inconvenientes de los métodos de recubrimiento convencionales (plasma spray, ablación láser...) cuya unión es por interacción mecánica, lo ...

  17. Photoluminescence of acupoint 'Waiqiu' in human superficial fascia

    International Nuclear Information System (INIS)

    Zhang Yuan; Yan Xiaohui; Liu Chenglin; Dang Ruishan; Zhang Xinyi

    2006-01-01

    The spectral characters of an acupuncture point named 'Waiqiu' in superficial fascia tissue have been studied by photoluminescence (PL) spectroscopy under the excitation of 457.9 nm. The PL around 'Waiqiu' acupuncture point consists of two sub-bands resulting from the flavin adenine dinucleotide (FAD) and phospholipids, and the porphyrins (including purine, isoxanthopterin and tryptophan), respectively. More emission due to FAD and phospholipids is found inside the acupuncture effect area of 'Waiqiu' than its marginal or outside acupuncture regions. The ratio of emission intensity of FAD and phospholipids to one of porphyrins gradually decreases along the direction away from the center of the acupuncture point. It implies that the component proportion changes between FAD, phospholipids and porphyrins around the 'Waiqiu' acupuncture point. We suggest that there might be a certain relationship between redox function of FAD and 'Waiqiu' acupuncture effect

  18. Transient superficial peroneal nerve palsy after anterior cruciate ligament reconstruction

    Directory of Open Access Journals (Sweden)

    Majed Alrowaili

    2016-06-01

    Full Text Available A 19-year-old male subject was diagnosed with medial meniscal, lateral meniscal and anterior cruciate ligament (ACL tear. The symptoms did not subside after 4 months of physical therapy, and he underwent arthroscopic partial medial and lateral meniscectomy and ACL reconstruction. Immediately after the patient woke up from general anesthesia, he started experience loss of sensation in the area of superficial peroneal nerve with inverted dorsiflexion of foot and ankle. Instantly, the bandage and knee brace was removed and a diagnosis of compartment syndrome was ruled out. After eight hours, post-operatively, the patient started receiving physiotherapy. He complained of numbness and tingling in the same area. After 24 h, post-operatively, the patient started to regain dorsiflexion and eversion gradually. Two days after the surgery, the patient exhibited complete recovery of neurological status.

  19. Superficial Granulomatous Pyoderma Gangrenosum of the Penis: A Case Report

    Directory of Open Access Journals (Sweden)

    Shyamala S. Gopi

    2006-01-01

    Full Text Available Classic type of pyoderma gangrenosum (PG is an uncommon ulceronecrotic cutaneous disease of uncertain aetiology characterised by broad zones of confluent ulceration with violaceous undermined margins. Some 50% of cases are associated with systemic diseases. The superficial granulomatous variant of pyoderma gangrenosum (SGPG of the external genitalia is extremely rare Patients with this condition develop single or multiple ulcerated skin lesions often with sinus tract formation. The majority of these lesions were found on the trunk and limbs. SGPG is less likely to be associated with underlying disease processes than classic PG. We present a 58 year-old with recalcitrant penile ulceration demonstrated to be SGPG on biopsy. Although rare and poorly recognised, the histological features are sufficiently typical to allow the correct diagnosis to be established.

  20. Feedback Interactions of Polymerized Actin with the Cell Membrane: Waves, Pulses, and Oscillations

    Science.gov (United States)

    Carlsson, Anders

    Polymerized filaments of the protein actin have crucial functions in cell migration, and in bending the cell membrane to drive endocytosis or the formation of protrusions. The nucleation and polymerization of actin filaments are controlled by upstream agents in the cell membrane, including nucleation-promoting factors (NPFs) that activate the Arp2/3 complex to form new branches on pre-existing filaments. But polymerized actin (F-actin) also feeds back on the assembly of NPFs. We explore the effects of the resulting feedback loop of F-actin and NPFs on two phenomena: actin pulses that drive endocytosis in yeast, and actin waves traveling along the membrane of several cell types. In our model of endocytosis in yeast, the actin network is grown explicitly in three dimensions, exerts a negative feedback interaction on localized patch of NPFs in the membrane, and bends the membrane by exerting a distribution of forces. This model explains observed actin and NPF pulse dynamics, and the effects of several interventions including i) NPF mutations, ii) inhibition of actin polymerization, and iii) deletion of a protein that allows F-actin to bend the cell membrane. The model predicts that mutation of the active region of an NPF will enhance the accumulation of that NPF, and we confirm this prediction by quantitative fluorescence microscopy. For actin waves, we treat a similar model, with NPFs distributed over a larger region of the cell membrane. This model naturally generates actin waves, and predicts a transition from wave behavior to spatially localized oscillations when NPFs are confined to a small region. We also predict a transition from waves to static polarization as the negative-feedback coupling between F-actin and the NPFs is reduced. Supported by NIGMS Grant R01 GM107667.

  1. Superficial plantar cutaneous sensation does not trigger barefoot running adaptations.

    Science.gov (United States)

    Thompson, M A; Hoffman, K M

    2017-09-01

    It has long been proposed that the gait alterations associated with barefoot running are mediated by alterations in sensory feedback, yet there has been no data to support this claim. Thus, the purpose of this study was to examine the role of superficial plantar cutaneous feedback in barefoot and shod running. 10 healthy active subjects (6 male, 4 female); mass: 65.2+9.7kg; age: 27+7.1years participated in this study. 10 over-ground running trials were completed in each of the following conditions: barefoot (BF), shod (SHOD), anesthetized barefoot (ANEST BF) and anesthetized shod (ANEST SHOD). For the anesthetized conditions 0.1-0.3mL of 1% lidocaine was injected into the dermal layer of the plantar foot below the metatarsal heads, lateral column and heel. 3-dimensional motion analysis and ground reaction force (GRF) data were captured as subjects ran over a 20m runway with a force plate at 12m. Kinematic and kinetic differences were analyzed via two-way repeated measure ANOVAs. The differences in gait between the BF and SHOD conditions were consistent with previous research, with subjects exhibiting a significant decrease in stride length and changing from rearfoot strike when SHOD to fore/midfoot strike when BF. Additionally, BF running was associated with decreased impact peak magnitudes and peak vertical GRFs. Despite anesthetizing the plantar surface, there was no difference between the BF and ANEST BF conditions in terms of stride length, foot strike or GRFs. Superficial cutaneous sensory receptors are not primarily responsible for the gait changes associated with barefoot running. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Excimer laser superficial keratectomy for proud nebulae in keratoconus.

    Science.gov (United States)

    Moodaley, L; Liu, C; Woodward, E G; O'Brart, D; Muir, M K; Buckley, R

    1994-06-01

    Contact lens intolerance in keratoconus may be due to the formation of a proud nebula at or near the apex of the cone. Excimer laser superficial keratectomy was performed as an outpatients with proud nebulae as treatment patients with proud nebulae as treatment for their contact lens intolerance. The mean period of contact lens wear before the development of intolerance was 13.4 years (range 2 to 27 years). Following the development of intolerance, three patients abandoned contact lens wear in the affected eye while the remainder experienced a reduction in comfortable wearing time (mean = 3.75 hours; range: 0-14 hours). All patients had good potential Snellen visual acuity with a contact lens of 6/9 (nine eyes) and 6/12 (one eye). The proud nebulae were directly ablated with a 193 nm ArF excimer laser using a 1 mm diameter beam. Between 100-150 pulses were sufficient to ablate the raised area. Patients experienced no pain during the procedure and reported minimal discomfort postoperatively. In all cases flattening of the proud nebulae was achieved. Seven patients were able to resume regular contact lens wear (mean wearing time = 10.17 hours; range 8 to 16 hours). In three patients, resumption of contact lens wear was unsuccessful because of cone steepness. All patients achieved postoperative Snellen visual acuity of 6/12 or better with a contact lens. Four patients experienced a loss of one line in Snellen acuity. The mean follow up period was 8.3 months (range 2 to 17 months). Excimer laser superficial keratectomy is a useful technique for the treatment of contact lens intolerance caused by proud nebulae in patients with keratoconus. Penetrating keratoplasty is thus avoided.

  3. Plant actin cytoskeleton re-modeling by plant parasitic nematodes.

    Science.gov (United States)

    Engler, Janice de Almeida; Rodiuc, Natalia; Smertenko, Andrei; Abad, Pierre

    2010-03-01

    The cytoskeleton is an important component of the plant's defense mechanism against the attack of pathogenic organisms. Plants however, are defenseless against parasitic root-knot and cyst nematodes and respond to the invasion by the development of a special feeding site that supplies the parasite with nutrients required for the completion of its life cycle. Recent studies of nematode invasion under treatment with cytoskeletal drugs and in mutant plants where normal functions of the cytoskeleton have been affected, demonstrate the importance of the cytoskeleton in the establishment of a feeding site and successful nematode reproduction. It appears that in the case of microfilaments, nematodes hijack the intracellular machinery that regulates actin dynamics and modulate the organization and properties of the actin filament network. Intervening with this process reduces the nematode infection efficiency and inhibits its life cycle. This discovery uncovers a new pathway that can be exploited for the protection of plants against nematodes.

  4. Prokaryotic DNA segregation by an actin-like filament

    DEFF Research Database (Denmark)

    Møller-Jensen, Jakob; Bugge Jensen, Rasmus; Löwe, Jan

    2002-01-01

    The mechanisms responsible for prokaryotic DNA segregation are largely unknown. The partitioning locus (par) encoded by the Escherichia coli plasmid R1 actively segregates its replicon to daughter cells. We show here that the ParM ATPase encoded by par forms dynamic actin-like filaments with prop...... point for ParM polymerization. Hence, we provide evidence for a simple prokaryotic analogue of the eukaryotic mitotic spindle apparatus.......The mechanisms responsible for prokaryotic DNA segregation are largely unknown. The partitioning locus (par) encoded by the Escherichia coli plasmid R1 actively segregates its replicon to daughter cells. We show here that the ParM ATPase encoded by par forms dynamic actin-like filaments...

  5. Actin depolymerization enhances adipogenic differentiation in human stromal stem cells

    DEFF Research Database (Denmark)

    Chen, Li; Hu, Huimin; Qiu, Weimin

    2018-01-01

    Human stromal stem cells (hMSCs) differentiate into adipocytes that play a role in skeletal tissue homeostasis and whole body energy metabolism. During adipocyte differentiation, hMSCs exhibit significant changes in cell morphology suggesting changes in cytoskeletal organization. Here, we examined...... differentiation as evidenced by decreased number of mature adipocytes and decreased adipocyte specific gene expression (ADIPOQ, LPL, PPARG, FABP4). In contrast, disruption of actin cytoskeleton by Cytochalasin D enhanced adipocyte differentiation. Follow up studies revealed that the effects of CFL1 on adipocyte...... differentiation depended on the activity of LIM domain kinase 1 (LIMK1) which is the major upstream kinase of CFL1. Inhibiting LIMK by its specific chemical inhibitor LIMKi inhibited the phosphorylation of CFL1 and actin polymerization, and enhanced the adipocyte differentiation. Moreover, treating h...

  6. Health related quality of life in patients with actinic keratosis

    DEFF Research Database (Denmark)

    Tennvall, Gunnel Ragnarson; Norlin, J M; Malmberg, I

    2015-01-01

    BACKGROUND: Actinic keratosis (AK) is a common skin condition that may progress to non-melanoma skin cancer (NMSC). The disease may influence Health Related Quality of Life (HRQoL), but studies of HRQoL in patients with AK are limited. The purpose of the study was to analyze HRQoL in patients......-center setting. Dermatologists assessed AK severity and patients completed: Actinic Keratosis Quality of Life Questionnaire (AKQoL), Dermatology Life Quality Index (DLQI), and EQ-5D-5 L including EQ-VAS. Differences between categorical subgroups were tested with Wilcoxon rank-sum test. The relationship between...... with different severity levels of AK treated in dermatology specialist care using generic and disease-specific HRQoL instruments and to analyze their relationship. METHODS: AK patients who visited dermatological clinics in Denmark were included in an observational, cross-sectional, study in a multi...

  7. Mapping of the Mouse Actin Capping Protein Beta Subunit Gene

    Directory of Open Access Journals (Sweden)

    Cooper John A

    2000-07-01

    Full Text Available Abstract Background Capping protein (CP, a heterodimer of α and β subunits, is found in all eukaryotes. CP binds to the barbed ends of actin filaments in vitro and controls actin assembly and cell motility in vivo. Vertebrates have three isoforms of CPβ produced by alternatively splicing from one gene; lower organisms have one gene and one isoform. Results We isolated genomic clones corresponding to the β subunit of mouse CP and identified its chromosomal location by interspecies backcross mapping. Conclusions The CPβ gene (Cappb1 mapped to Chromosome 4 between Cdc42 and D4Mit312. Three mouse mutations, snubnose, curly tail, and cribriform degeneration, map in the vicinity of the β gene.

  8. Osmosensation in vasopressin neurons: changing actin density to optimize function.

    Science.gov (United States)

    Prager-Khoutorsky, Masha; Bourque, Charles W

    2010-02-01

    The proportional relation between circulating vasopressin concentration and plasma osmolality is fundamental for body fluid homeostasis. Although changes in the sensitivity of this relation are associated with pathophysiological conditions, central mechanisms modulating osmoregulatory gain are unknown. Here, we review recent data that sheds important light on this process. The cell autonomous osmosensitivity of vasopressin neurons depends on cation channels comprising a variant of the transient receptor potential vanilloid 1 (TRPV1) channel. Hyperosmotic activation is mediated by a mechanical process where sensitivity increases in proportion with actin filament density. Moreover, angiotensin II amplifies osmotic activation by a rapid stimulation of actin polymerization, suggesting that neurotransmitter-induced changes in cytoskeletal organization in osmosensory neurons can mediate central changes in osmoregulatory gain. (c) 2009 Elsevier Ltd. All rights reserved.

  9. Actinic inspection of multilayer defects on EUV masks

    International Nuclear Information System (INIS)

    Barty, A; Liu, Y; Gullikson, E; Taylor, J S; Wood, O

    2005-01-01

    The production of defect-free mask blanks, and the development of techniques for inspecting and qualifying EUV mask blanks, remains a key challenge for EUV lithography. In order to ensure a reliable supply of defect-free mask blanks, it is necessary to develop techniques to reliably and accurately detect defects on un-patterned mask blanks. These inspection tools must be able to accurately detect all critical defects whilst simultaneously having the minimum possible false-positive detection rate. There continues to be improvement in high-speed non-actinic mask blank inspection tools, and it is anticipated that these tools can and will be used by industry to qualify EUV mask blanks. However, the outstanding question remains one of validating that non-actinic inspection techniques are capable of detecting all printable EUV defects. To qualify the performance of non-actinic inspection tools, a unique dual-mode EUV mask inspection system has been installed at the Advanced Light Source (ALS) synchrotron at Lawrence Berkeley National Laboratory. In high-speed inspection mode, whole mask blanks are scanned for defects using 13.5-nm wavelength light to identify and map all locations on the mask that scatter a significant amount of EUV light. In imaging, or defect review mode, a zone plate is placed in the reflected beam path to image a region of interest onto a CCD detector with an effective resolution on the mask of 100-nm or better. Combining the capabilities of the two inspection tools into one system provides the unique capability to determine the coordinates of native defects that can be used to compare actinic defect inspection with visible light defect inspection tools under commercial development, and to provide data for comparing scattering models for EUV mask defects

  10. Oral acetylsalicylic acid and prevalence of actinic keratosis.

    Science.gov (United States)

    Schmitt, Juliano; Miot, Hélio

    2014-01-01

    To investigate the influence of a regular oral use of acetylsalicylic acid in the prevalence of actinic keratosis. A case-control study with dermatologic outpatients above 50 years of age assessed between 2009 and 2011. Cases were defined as those who had been under regular use of oral acetylsalicylic acid for more than six consecutive months. The assessment focused on: age, sex, skin-type, tobacco smoking, use of medication, occurrence of individual or family skin cancer, and sunscreen and sun exposure habits. Actinic keratoses were counted in the medial region of the face and upper limbs. Counts were adjusted by co-variables based on a generalized linear model. A total of 74 cases and 216 controls were assessed. The median time of acetylsalicylic acid use was 36 months. Cases differed from controls as to the highest age, highest prevalence of use of angiotensin-converting enzyme inhibitors and fewer keratosis on the face and on the upper limbs (pkeratosis and upper-limb erythematous actinic keratosis (pkeratosis, especially facial and erythematous ones.

  11. New Topical Treatment Options for Actinic Keratosis: A Systematic Review.

    Science.gov (United States)

    Stockfleth, Eggert; Sibbring, Gillian C; Alarcon, Ivette

    2016-01-01

    This systematic review compared the relative efficacy of 5-fluorouracil 0.5% in salicylic acid 10% (5-FU/SA), ingenol mebutate (IMB) and imiquimod 2.5%/3.75% (IMI) for actinic keratosis on the face, forehead or scalp. Only 11 publications, relating to 7 randomised controlled trials, met inclusion criteria and it was only possible to compare the effect of all 3 treatments on complete clinical clearance, and the effect of 5-FU/SA and IMB on actinic keratosis recurrence rate. Despite a higher vehicle response rate for 5-FU/SA, complete clinical clearance was higher than IMB and IMI (55.4, 42.2, and 25.0-30.6/34.0-35.6%, [corrected] respectively). 5-FU/SA was also associated with lower actinic keratosis recurrence rate than IMB at 12 months post-treatment (32.7 vs. 53.9%). Although qualitative assessment suggested a numerical advantage of 5-FU/SA over IMB and IMI in terms of complete clinical clearance and sustained clearance, clinical data from longer term trials, with comparable outcome measures, are required to corroborate these findings.

  12. Antibodies to filamentous actin (F-actin) in type 1 autoimmune hepatitis.

    Science.gov (United States)

    Granito, A; Muratori, L; Muratori, P; Pappas, G; Guidi, M; Cassani, F; Volta, U; Ferri, A; Lenzi, M; Bianchi, F B

    2006-03-01

    To evaluate the diagnostic significance of anti-filamentous actin antibodies (A-FAA) assessed with a commercial ELISA in comparison with immunofluorescence reactivity and patterns of anti-smooth muscle antibodies (SMA); and to correlate A-FAA positivity with clinical, immunogenetic, laboratory, and histological features in patients with autoimmune hepatitis type 1 (AIH-1). We studied 78 consecutive untreated AIH-1 patients and 160 controls: 22 with autoimmune hepatitis type 2 (AIH-2), 51 with hepatitis C, 17 with coeliac disease (CD), 20 with primary biliary cirrhosis (PBC) and 50 blood donors. SMA was evaluated by indirect immunofluorescence (IIF) on frozen sections of rat tissues, and A-FAA with a modified commercial ELISA. SMA was detected by IIF in 61 (78%) of 78 AIH-1 patients, of whom 47 (60%) had the SMA-T/G and 14 (18%) the SMA-V pattern. Of the pathological controls, 32 (20%) had the SMA-V pattern (25 with hepatitis C, 2 with AIH-2, 2 with PBC, 3 with CD). A-FAA were present in 55 AIH-1 patients (70.5%; 46 with SMA-T/G, 7 with SMA-V, and 2 SMA-negative), and in 10 controls (6%), of whom five had hepatitis C, two AIH-2, two PBC and one CD. The association between A-FAA and the SMA-T/G pattern was statistically significant (p<0.0001). A-FAA levels were higher in SMA-T/G positive than SMA-V positive AIH-1 patients and controls (p<0.0001). A-FAA positivity was significantly associated with higher gamma-globulin and IgG levels, but did not correlate with other considered parameters. The modified A-FAA ELISA strictly correlates with the SMA-T/G pattern and is a reliable and operator independent assay for AIH-1. Detection of A-FAA, even if devoid of prognostic relevance, may be useful when interpretative doubts of standard IIF arise.

  13. Antibodies to filamentous actin (F‐actin) in type 1 autoimmune hepatitis

    Science.gov (United States)

    Granito, A; Muratori, L; Muratori, P; Pappas, G; Guidi, M; Cassani, F; Volta, U; Ferri, A; Lenzi, M; Bianchi, F B

    2006-01-01

    Aims To evaluate the diagnostic significance of anti‐filamentous actin antibodies (A‐FAA) assessed with a commercial ELISA in comparison with immunofluorescence reactivity and patterns of anti‐smooth muscle antibodies (SMA); and to correlate A‐FAA positivity with clinical, immunogenetic, laboratory, and histological features in patients with autoimmune hepatitis type 1 (AIH‐1). Methods We studied 78 consecutive untreated AIH‐1 patients and 160 controls: 22 with autoimmune hepatitis type 2 (AIH‐2), 51 with hepatitis C, 17 with coeliac disease (CD), 20 with primary biliary cirrhosis (PBC) and 50 blood donors. SMA was evaluated by indirect immunofluorescence (IIF) on frozen sections of rat tissues, and A‐FAA with a modified commercial ELISA. Results SMA was detected by IIF in 61 (78%) of 78 AIH‐1 patients, of whom 47 (60%) had the SMA‐T/G and 14 (18%) the SMA‐V pattern. Of the pathological controls, 32 (20%) had the SMA‐V pattern (25 with hepatitis C, 2 with AIH‐2, 2 with PBC, 3 with CD). A‐FAA were present in 55 AIH‐1 patients (70.5%; 46 with SMA‐T/G, 7 with SMA‐V, and 2 SMA‐negative), and in 10 controls (6%), of whom five had hepatitis C, two AIH‐2, two PBC and one CD. The association between A‐FAA and the SMA‐T/G pattern was statistically significant (p<0.0001). A‐FAA levels were higher in SMA‐T/G positive than SMA‐V positive AIH‐1 patients and controls (p<0.0001). A‐FAA positivity was significantly associated with higher γ‐globulin and IgG levels, but did not correlate with other considered parameters. Conclusion The modified A‐FAA ELISA strictly correlates with the SMA‐T/G pattern and is a reliable and operator independent assay for AIH‐1. Detection of A‐FAA, even if devoid of prognostic relevance, may be useful when interpretative doubts of standard IIF arise. PMID:16505279

  14. Altered Cell Mechanics from the Inside: Dispersed Single Wall Carbon Nanotubes Integrate with and Restructure Actin

    Directory of Open Access Journals (Sweden)

    Mohammad F. Islam

    2012-05-01

    Full Text Available With a range of desirable mechanical and optical properties, single wall carbon nanotubes (SWCNTs are a promising material for nanobiotechnologies. SWCNTs also have potential as biomaterials for modulation of cellular structures. Previously, we showed that highly purified, dispersed SWCNTs grossly alter F-actin inside cells. F-actin plays critical roles in the maintenance of cell structure, force transduction, transport and cytokinesis. Thus, quantification of SWCNT-actin interactions ranging from molecular, sub-cellular and cellular levels with both structure and function is critical for developing SWCNT-based biotechnologies. Further, this interaction can be exploited, using SWCNTs as a unique actin-altering material. Here, we utilized molecular dynamics simulations to explore the interactions of SWCNTs with actin filaments. Fluorescence lifetime imaging microscopy confirmed that SWCNTs were located within ~5 nm of F-actin in cells but did not interact with G-actin. SWCNTs did not alter myosin II sub-cellular localization, and SWCNT treatment in cells led to significantly shorter actin filaments. Functionally, cells with internalized SWCNTs had greatly reduced cell traction force. Combined, these results demonstrate direct, specific SWCNT alteration of F-actin structures which can be exploited for SWCNT-based biotechnologies and utilized as a new method to probe fundamental actin-related cellular processes and biophysics.

  15. Gamma interferon-induced guanylate binding protein 1 is a novel actin cytoskeleton remodeling factor.

    Science.gov (United States)

    Ostler, Nicole; Britzen-Laurent, Nathalie; Liebl, Andrea; Naschberger, Elisabeth; Lochnit, Günter; Ostler, Markus; Forster, Florian; Kunzelmann, Peter; Ince, Semra; Supper, Verena; Praefcke, Gerrit J K; Schubert, Dirk W; Stockinger, Hannes; Herrmann, Christian; Stürzl, Michael

    2014-01-01

    Gamma interferon (IFN-γ) regulates immune defenses against viruses, intracellular pathogens, and tumors by modulating cell proliferation, migration, invasion, and vesicle trafficking processes. The large GTPase guanylate binding protein 1 (GBP-1) is among the cellular proteins that is the most abundantly induced by IFN-γ and mediates its cell biologic effects. As yet, the molecular mechanisms of action of GBP-1 remain unknown. Applying an interaction proteomics approach, we identified actin as a strong and specific binding partner of GBP-1. Furthermore, GBP-1 colocalized with actin at the subcellular level and was both necessary and sufficient for the extensive remodeling of the fibrous actin structure observed in IFN-γ-exposed cells. These effects were dependent on the oligomerization and the GTPase activity of GBP-1. Purified GBP-1 and actin bound to each other, and this interaction was sufficient to impair the formation of actin filaments in vitro, as demonstrated by atomic force microscopy, dynamic light scattering, and fluorescence-monitored polymerization. Cosedimentation and band shift analyses demonstrated that GBP-1 binds robustly to globular actin and slightly to filamentous actin. This indicated that GBP-1 may induce actin remodeling via globular actin sequestering and/or filament capping. These results establish GBP-1 as a novel member within the family of actin-remodeling proteins specifically mediating IFN-γ-dependent defense strategies.

  16. Mechanical coupling between transsynaptic N-cadherin adhesions and actin flow stabilizes dendritic spines

    Science.gov (United States)

    Chazeau, Anaël; Garcia, Mikael; Czöndör, Katalin; Perrais, David; Tessier, Béatrice; Giannone, Grégory; Thoumine, Olivier

    2015-01-01

    The morphology of neuronal dendritic spines is a critical indicator of synaptic function. It is regulated by several factors, including the intracellular actin/myosin cytoskeleton and transcellular N-cadherin adhesions. To examine the mechanical relationship between these molecular components, we performed quantitative live-imaging experiments in primary hippocampal neurons. We found that actin turnover and structural motility were lower in dendritic spines than in immature filopodia and increased upon expression of a nonadhesive N-cadherin mutant, resulting in an inverse relationship between spine motility and actin enrichment. Furthermore, the pharmacological stimulation of myosin II induced the rearward motion of actin structures in spines, showing that myosin II exerts tension on the actin network. Strikingly, the formation of stable, spine-like structures enriched in actin was induced at contacts between dendritic filopodia and N-cadherin–coated beads or micropatterns. Finally, computer simulations of actin dynamics mimicked various experimental conditions, pointing to the actin flow rate as an important parameter controlling actin enrichment in dendritic spines. Together these data demonstrate that a clutch-like mechanism between N-cadherin adhesions and the actin flow underlies the stabilization of dendritic filopodia into mature spines, a mechanism that may have important implications in synapse initiation, maturation, and plasticity in the developing brain. PMID:25568337

  17. FIMBRIN1 is involved in lily pollen tube growth by stabilizing the actin fringe.

    Science.gov (United States)

    Su, Hui; Zhu, Jinsheng; Cai, Chao; Pei, Weike; Wang, Jiaojiao; Dong, Huaijian; Ren, Haiyun

    2012-11-01

    An actin fringe structure in the subapex plays an important role in pollen tube tip growth. However, the precise mechanism by which the actin fringe is generated and maintained remains largely unknown. Here, we cloned a 2606-bp full-length cDNA encoding a deduced 77-kD fimbrin-like protein from lily (Lilium longiflorum), named FIMBRIN1 (FIM1). Ll-FIM1 was preferentially expressed in pollen and concentrated at actin fringe in the subapical region, as well as in longitudinal actin-filament bundles in the shank of pollen tubes. Microinjection of Ll-FIM1 antibody into lily pollen tubes inhibited tip growth and disrupted the actin fringe. Furthermore, we verified the function of Ll-FIM1 in the fim5 mutant of its closest relative, Arabidopsis thaliana. Pollen tubes of fim5 mutants grew with a larger diameter in early stages but could recover into normal forms in later stages, despite significantly slower growth rates. The actin fringe of the fim5 mutants, however, was impaired during both early and late stages. Impressively, stable expression of fim5pro:GFP:Ll-FIM1 rescued the actin fringe and the growth rate of Arabidopsis fim5 pollen tubes. In vitro biochemical analysis showed that Ll-FIM1 could bundle actin filaments. Thus, our study has identified a fimbrin that may stabilize the actin fringe by cross-linking actin filaments into bundles, which is important for proper tip growth of lily pollen tubes.

  18. WICH, a member of WASP-interacting protein family, cross-links actin filaments

    International Nuclear Information System (INIS)

    Kato, Masayoshi; Takenawa, Tadaomi

    2005-01-01

    In yeast, Verprolin plays an important role in rearrangement of the actin cytoskeleton. There are three mammalian homologues of Verprolin, WIP, CR16, and WICH, and all of them bind actin and Wiskott-Aldrich syndrome protein (WASP) and/or neural-WASP. Here, we describe a novel function of WICH. In vitro co-sedimentation analysis revealed that WICH not only binds to actin filaments but also cross-links them. Fluorescence and electron microscopy detected that this cross-linking results in straight bundled actin filaments. Overexpression of WICH alone in cultured fibroblast caused the formation of thick actin fibers. This ability of WICH depended on its own actin cross-linking activity. Importantly, the actin cross-linking activity of WICH was modified through a direct association with N-WASP. Taken together, these data suggest that WICH induces a bundled form of actin filament with actin cross-linking activity and the association with N-WASP suppresses that activity. WICH thus appears to be a novel actin bundling protein

  19. The actin cytoskeleton may control the polar distribution of an auxin transport protein

    Science.gov (United States)

    Muday, G. K.; Hu, S.; Brady, S. R.; Davies, E. (Principal Investigator)

    2000-01-01

    The gravitropic bending of plants has long been linked to the changes in the transport of the plant hormone auxin. To understand the mechanism by which gravity alters auxin movement, it is critical to know how polar auxin transport is initially established. In shoots, polar auxin transport is basipetal (i.e., from the shoot apex toward the base). It is driven by the basal localization of the auxin efflux carrier complex. One mechanism for localizing this efflux carrier complex to the basal membrane may be through attachment to the actin cytoskeleton. The efflux carrier protein complex is believed to consist of several polypeptides, including a regulatory subunit that binds auxin transport inhibitors, such as naphthylphthalamic acid (NPA). Several lines of experimentation have been used to determine if the NPA binding protein interacts with actin filaments. The NPA binding protein has been shown to partition with the actin cytoskeleton during detergent extraction. Agents that specifically alter the polymerization state of the actin cytoskeleton change the amount of NPA binding protein and actin recovered in these cytoskeletal pellets. Actin-affinity columns were prepared with polymers of actin purified from zucchini hypocotyl tissue. NPA binding activity was eluted in a single peak from the actin filament column. Cytochalasin D, which fragments the actin cytoskeleton, was shown to reduce polar auxin transport in zucchini hypocotyls. The interaction of the NPA binding protein with the actin cytoskeleton may localize it in one plane of the plasma membrane, and thereby control the polarity of auxin transport.

  20. Binding and assembly of actin filaments by plasma membranes from dictyostelium discoideum

    International Nuclear Information System (INIS)

    Schwartz, M.A.; Luna, E.J.

    1986-01-01

    The binding of native, 125 I-Bolton-Hunter-labeled actin to purified Dictyostelium discoideum plasma membranes was measured using a sedimentation assay. Binding was saturable only in the presence of the actin capping protein, gelsolin. The binding curves were sigmoidal, indicating positive cooperativity at low actin concentrations. This cooperativity appeared to be due to actin-actin associations during polymerization, since phalloidin converted the curve to a hyperbolic shape. This membrane-bound actin stained with rhodamine-phalloidin and was cross-linked by m-maleimidobenzoyl succinimide ester, a bifunctional cross-linker, into multimers with the same pattern observed for cross-linked F-actin. The authors conclude that D. discoideum plasma membranes bind actin specifically and saturably and that these membranes organize actin into filaments below the normal critical concentration for polymerization. This interaction probably occurs between multiple binding sites on the membrane and the side of the actin filament, and may be related to the clustering of membrane proteins

  1. Actin Nemaline Myopathy Mouse Reproduces Disease, Suggests Other Actin Disease Phenotypes and Provides Cautionary Note on Muscle Transgene Expression

    Science.gov (United States)

    Ravenscroft, Gianina; Jackaman, Connie; Sewry, Caroline A.; McNamara, Elyshia; Squire, Sarah E.; Potter, Allyson C.; Papadimitriou, John; Griffiths, Lisa M.; Bakker, Anthony J.; Davies, Kay E.; Laing, Nigel G.; Nowak, Kristen J.

    2011-01-01

    Mutations in the skeletal muscle α-actin gene (ACTA1) cause congenital myopathies including nemaline myopathy, actin aggregate myopathy and rod-core disease. The majority of patients with ACTA1 mutations have severe hypotonia and do not survive beyond the age of one. A transgenic mouse model was generated expressing an autosomal dominant mutant (D286G) of ACTA1 (identified in a severe nemaline myopathy patient) fused with EGFP. Nemaline bodies were observed in multiple skeletal muscles, with serial sections showing these correlated to aggregates of the mutant skeletal muscle α-actin-EGFP. Isolated extensor digitorum longus and soleus muscles were significantly weaker than wild-type (WT) muscle at 4 weeks of age, coinciding with the peak in structural lesions. These 4 week-old mice were ∼30% less active on voluntary running wheels than WT mice. The α-actin-EGFP protein clearly demonstrated that the transgene was expressed equally in all myosin heavy chain (MHC) fibre types during the early postnatal period, but subsequently became largely confined to MHCIIB fibres. Ringbinden fibres, internal nuclei and myofibrillar myopathy pathologies, not typical features in nemaline myopathy or patients with ACTA1 mutations, were frequently observed. Ringbinden were found in fast fibre predominant muscles of adult mice and were exclusively MHCIIB-positive fibres. Thus, this mouse model presents a reliable model for the investigation of the pathobiology of nemaline body formation and muscle weakness and for evaluation of potential therapeutic interventions. The occurrence of core-like regions, internal nuclei and ringbinden will allow analysis of the mechanisms underlying these lesions. The occurrence of ringbinden and features of myofibrillar myopathy in this mouse model of ACTA1 disease suggests that patients with these pathologies and no genetic explanation should be screened for ACTA1 mutations. PMID:22174871

  2. The structure of the actin-smooth muscle myosin motor domain complex in the rigor state.

    Science.gov (United States)

    Banerjee, Chaity; Hu, Zhongjun; Huang, Zhong; Warrington, J Anthony; Taylor, Dianne W; Trybus, Kathleen M; Lowey, Susan; Taylor, Kenneth A

    2017-12-01

    Myosin-based motility utilizes catalysis of ATP to drive the relative sliding of F-actin and myosin. The earliest detailed model based on cryo-electron microscopy (cryoEM) and X-ray crystallography postulated that higher actin affinity and lever arm movement were coupled to closure of a feature of the myosin head dubbed the actin-binding cleft. Several studies since then using crystallography of myosin-V and cryoEM structures of F-actin bound myosin-I, -II and -V have provided details of this model. The smooth muscle myosin II interaction with F-actin may differ from those for striated and non-muscle myosin II due in part to different lengths of important surface loops. Here we report a ∼6 Å resolution reconstruction of F-actin decorated with the nucleotide-free recombinant smooth muscle myosin-II motor domain (MD) from images recorded using a direct electron detector. Resolution is highest for F-actin and the actin-myosin interface (3.5-4 Å) and lowest (∼6-7 Å) for those parts of the MD at the highest radius. Atomic models built into the F-actin density are quite comparable to those previously reported for rabbit muscle actin and show density from the bound ADP. The atomic model of the MD, is quite similar to a recently published structure of vertebrate non-muscle myosin II bound to F-actin and a crystal structure of nucleotide free myosin-V. Larger differences are observed when compared to the cryoEM structure of F-actin decorated with rabbit skeletal muscle myosin subfragment 1. The differences suggest less closure of the 50 kDa domain in the actin bound skeletal muscle myosin structure. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. A peek into tropomyosin binding and unfolding on the actin filament.

    Directory of Open Access Journals (Sweden)

    Abhishek Singh

    Full Text Available BACKGROUND: Tropomyosin is a prototypical coiled coil along its length with subtle variations in structure that allow interactions with actin and other proteins. Actin binding globally stabilizes tropomyosin. Tropomyosin-actin interaction occurs periodically along the length of tropomyosin. However, it is not well understood how tropomyosin binds actin. PRINCIPAL FINDINGS: Tropomyosin's periodic binding sites make differential contributions to two components of actin binding, cooperativity and affinity, and can be classified as primary or secondary sites. We show through mutagenesis and analysis of recombinant striated muscle alpha-tropomyosins that primary actin binding sites have a destabilizing coiled-coil interface, typically alanine-rich, embedded within a non-interface recognition sequence. Introduction of an Ala cluster in place of the native, more stable interface in period 2 and/or period 3 sites (of seven increased the affinity or cooperativity of actin binding, analysed by cosedimentation and differential scanning calorimetry. Replacement of period 3 with period 5 sequence, an unstable region of known importance for cooperative actin binding, increased the cooperativity of binding. Introduction of the fluorescent probe, pyrene, near the mutation sites in periods 2 and 3 reported local instability, stabilization by actin binding, and local unfolding before or coincident with dissociation from actin (measured using light scattering, and chain dissociation (analyzed using circular dichroism. CONCLUSIONS: This, and previous work, suggests that regions of tropomyosin involved in binding actin have non-interface residues specific for interaction with actin and an unstable interface that is locally stabilized upon binding. The destabilized interface allows residues on the coiled-coil surface to obtain an optimal conformation for interaction with actin by increasing the number of local substates that the side chains can sample. We suggest

  4. The 5’cap of Tobacco Mosaic Virus (TMV) is required for virion attachment to the actin/ER network during early infection

    DEFF Research Database (Denmark)

    Christensen, Nynne Meyn; Tilsner, Jens; Bell, Karen

    to the motile cortical actin/ER network within minutes of injection. Granule movement on actin/ER was arrested by actin inhibitors indicating actindependent RNA movement. The 5’ methylguanosine TMV cap was shown to be required for vRNA anchoring to the ER. TMV vRNA lacking the 5’cap failed to form granules...... the fluorescent vRNA pool nor co-injected GFP left the injected trichome, indicating that the synthesis of unlabelled progeny viral (v)RNA is required to initiate cell-cell movement, and that virus movement is not accompanied by passive plasmodesmatal gating. Cy3-vRNA formed granules that became anchored...... on the same ER-bound granules, indicating that TMV virions may become attached to the ER prior to uncoating of the viral genome....

  5. Colonisation of basal cell carcinoma and actinic keratosis by malignant melanoma in situ in a patient with xeroderma pigmentosum variant

    Directory of Open Access Journals (Sweden)

    Louise J. Smith

    2012-04-01

    Full Text Available Although malignant melanoma (MM and both basal cell carcinoma (BCC and actinic keratosis (AK are sun-induced lesions, the coexistence of these entities at the same anatomical site (collision tumour is exceedingly rare. We report the case of a 54-year-old woman with a known history of xeroderma pigmentosum variant (XPV who presented with 2 separate skin lesions over the middle and upper right forearm, respectively. The clinical impression was that of BCCs or squamous cell lesions. On histological examination, both specimens showed features of melanoma in situ (MIS. In the first lesion, MIS merged with and colonised a superficial and focally invasive BCC. In the second lesion, MIS merged with an AK. No separate invasive nests of malignant melanoma were seen in either specimen. The atypical melanocytes were highlighted by Melan-A and HMB-45 immunostaining, whereas the epithelial cells in both the BCC and AK stained with the pancytokeratin MNF-116. The patient had a previous history of multiple MMs and non-melanomatous skin cancers and finally developed widespread metastatic malignant melanoma, which proved fatal. The rare and interesting phenomenon of collision tumours may pose diagnostic difficulties. To our knowledge, this is the first reported simultaneous presentation of cytologically malignant collision tumours in a patient with XPV.

  6. Medicina regenerativa y superficie ocular Regenerative medicine and ocular surface

    Directory of Open Access Journals (Sweden)

    Taimi Cárdenas Díaz

    2012-06-01

    Full Text Available En los últimos años se ha producido un extraordinario avance en los conocimientos relacionados con diferentes ramas biomédicas, entre ellas, la biología celular. Esto ha dado un notable impulso a una nueva rama de la medicina denominada medicina regenerativa. Esta nueva disciplina médica se basa fundamentalmente en los nuevos conocimientos sobre las células madre y en su capacidad de convertirse en células de diferentes tejidos. Una de las estructuras que tiene más interés, desde el punto de vista de la medicina regenerativa, es la superficie ocular. Por esto, se ha logrado notables progresos en la reconstrucción de la superficie ocular mediante la aplicación de procederes regenerativos. Los cultivos de células del epitelio corneal humano están siendo utilizados en el tratamiento de la insuficiencia limbar. Esto permite disminuir la incidencia de opacidades tipo Haze tras queratectomía fotorrefractiva, acelerar la curación y evitar la aparición de leucomas en pacientes con defectos epiteliares persistentes e intentar disminuir la incidencia de opacidades corneales en pacientes sometidos a queratectomía fototerapéutica por diferentes distrofias corneales.In the last few years, an extraordinary advance has taken place in the knowledge about several biomedical branches as is the case of cellular biology, which has remarkably encouraged the development of a new medical branch called regenerative medicine. This medical discipline is fundamentally based on the new knowledge on the stem cells and their capacity to become cells for different tissues. One of the most interesting structures for the regenerative medicine is the ocular surface. In the last few years, significant advances have been achieved in the field of the ocular surface reconstruction with regenerative procedures. Some cell cultures of the human corneal epithelium are being used to treat limber insufficiency, to reduce the incidence of haze-type opacities after

  7. 5-Aminolevulinic acid photodynamic therapy for superficial basal cell carcinoma

    International Nuclear Information System (INIS)

    Want, David; Kennedy, James C.; Brundage, Michael; Rothwell, Deanna

    1997-01-01

    Treatment of superficial basal cell carcinoma with topical 5-aminolevulinic acid photodynamic therapy (ALA-PDT) offers an alternative to plastic surgery and radiotherapy with potential for good cosmetic outcome and local control of disease. We report our clinical experience with this technique. Patients were treated prospectively on a study protocol enrolling a total of 118 patients (63 male, 55 female) with an average age of 65 years. Consecutive patients meeting eligibility criteria were invited to participate over a four year period. Median followup was 27 months (range 1 to 76 months). In the study group, 62 patients had single lesions and 56 had multiple lesions. Of the 56 patients with multiple lesions, 33 had 2-4 lesions, 11 had 5-9, and 11 had 10 or more. All patients were treated with 20% ALA dissolved in Glaxal Base applied to the tumors for three to four hours. Following removal of the cream, fluorescence intensity and distribution were assessed using a UV-A lamp, and the lesions were exposed to photoactivating light of wavelength greater than 600 nm for a light dose ranging from 100-150 J/cm2. Lesions were reassessed in followup, and scored as complete or partial responses. At subsequent patient assessments, lesions were scored as continued complete responses or recurrences. In the patients with single lesions, there was an initial complete response rate of 90.3%. Of the 56 patients with multiple lesions, 44 had all of their lesions respond completely, and there was an overall average response rate of 95.5%. Sixty three percent of males and 44% of females had all of their lesions respond completely. (p=0.033, Chi-squared test). There was no difference in response rate with respect to age, or site of lesion. The recurrence rates were 35% for patients with single lesions, and 10.5% for patients with multiple lesions. ALA-PDT would appear to be a promising alternative to conventional treatment for superficial basal cell carcinoma. Based on these results

  8. Inhibiting actin depolymerization enhances osteoblast differentiation and bone formation in human stromal stem cells

    DEFF Research Database (Denmark)

    Chen, Li; Shi, Kaikai; Frary, Charles

    2015-01-01

    Remodeling of the actin cytoskeleton through actin dynamics is involved in a number of biological processes, but its role in human stromal (skeletal) stem cells (hMSCs) differentiation is poorly understood. In the present study, we demonstrated that stabilizing actin filaments by inhibiting gene...... expression of the two main actin depolymerizing factors (ADFs): Cofilin 1 (CFL1) and Destrin (DSTN) in hMSCs, enhanced cell viability and differentiation into osteoblastic cells (OB) in vitro, as well as heterotopic bone formation in vivo. Similarly, treating hMSC with Phalloidin, which is known to stabilize...... polymerized actin filaments, increased hMSCs viability and OB differentiation. Conversely, Cytocholasin D, an inhibitor of actin polymerization, reduced cell viability and inhibited OB differentiation of hMSC. At a molecular level, preventing Cofilin phosphorylation through inhibition of LIM domain kinase 1...

  9. System-wide organization of actin cytoskeleton determines organelle transport in hypocotyl plant cells

    Science.gov (United States)

    Nowak, Jacqueline; Ivakov, Alexander; Somssich, Marc; Persson, Staffan; Nikoloski, Zoran

    2017-01-01

    The actin cytoskeleton is an essential intracellular filamentous structure that underpins cellular transport and cytoplasmic streaming in plant cells. However, the system-level properties of actin-based cellular trafficking remain tenuous, largely due to the inability to quantify key features of the actin cytoskeleton. Here, we developed an automated image-based, network-driven framework to accurately segment and quantify actin cytoskeletal structures and Golgi transport. We show that the actin cytoskeleton in both growing and elongated hypocotyl cells has structural properties facilitating efficient transport. Our findings suggest that the erratic movement of Golgi is a stable cellular phenomenon that might optimize distribution efficiency of cell material. Moreover, we demonstrate that Golgi transport in hypocotyl cells can be accurately predicted from the actin network topology alone. Thus, our framework provides quantitative evidence for system-wide coordination of cellular transport in plant cells and can be readily applied to investigate cytoskeletal organization and transport in other organisms. PMID:28655850

  10. Profilin as a regulator of the membrane-actin cytoskeleton interface in plant cells

    Directory of Open Access Journals (Sweden)

    Tiantian eSun

    2013-12-01

    Full Text Available Membrane structures and cytoskeleton dynamics are intimately inter-connected in the eukaryotic cell. Recently, the molecular mechanisms operating at this interface have been progressively addressed. Many experiments have revealed that the actin cytoskeleton can interact with membranes through various discrete membrane domains. The actin-binding protein, profilin has been proven to inhibit actin polymerization and to promote F-actin elongation. This is dependent on many factors, such as the profilin/G-actin ratio and the ionic environment of the cell. Additionally, profilin has specific domains that interact with phosphoinositides and poly-L-proline rich proteins; theoretically, this gives profilin the opportunity to interact with membranes, and a large number of experiments have confirmed this possibility. In this article, we summarize recent findings in plant cells, and discuss the evidence of the connections among actin cytoskeleton, profilin and biomembranes through direct or indirect relationships.

  11. Spinal meningeal melanocytoma and hydrocephalus and intracranial superficial siderosis

    International Nuclear Information System (INIS)

    Das, A.; Ratnagopal, P.; Puvanendran, K.; Teo, J.G.C.

    2001-01-01

    Meningeal melanocytomas are uncommon tumours of the central nervous system; fewer than 50 cases have been reported in the English literature. We review the unique clinical presentation, radiological appearance and histological features of a rare case of meningeal melanocytoma. The patient was a 50-year-old man with a history of hypertension who presented with complaints of severe headache, nausea, vomiting and blurry vision for 2 days. Clinical examination revealed no hyperpigmentation marks and he had no history of regressed skin melanocytic lesions. Apart from mild terminal neck stiffness, his general medical examination was unremarkable. On funduscopic examination he had marked bilateral papilloedema and the blind spots were enlarged bilaterally. His neurological examination was otherwise unremarkable. A magnetic resonance imaging scan (MRI) of the brain showed hydrocephalus. On the precontrast T 2 -weighted images, there was hyperintensity of the meninges with little change after administration of gadolinium, which was suggestive of blood. On the T 2 -weighted sequences, there was evidence of hypo-i intensity on the surface of the pons and medulla, which was indicative of superficial siderosis

  12. Current status of superficial pharyngeal squamous cell carcinoma in Japan.

    Science.gov (United States)

    Rikitake, Ryoko; Ando, Mizuo; Saito, Yuki; Yoshimoto, Seiichi; Yamasoba, Tatsuya; Higashi, Takahiro

    2017-10-01

    To investigate the status and treatment of superficial pharyngeal squamous cell carcinoma in Japan. We analyzed all cases diagnosed between 2011 and 2013, as recorded in the national database of hospital-based cancer registries. We extracted data on patient sex, age, tumor locations, histology, presentation routes, initial treatments, and TNM stages. Additionally, we compared the characteristics of pharyngeal carcinoma to those of esophageal cancer. A total of 16,521 oropharyngeal and hypopharyngeal cancers from 409 institutions were included. Diagnosis of Tis tumors was infrequent, and both cancers were likely to be diagnosed at an advanced stage (n = 866, 5.3%). Tis diseases were the most commonly detected during follow-up examinations for other diseases (n = 608, 70%). While more oropharyngeal Tis patients were men compared to T1-4 patients (88 vs 82%, respectively), hypopharyngeal cancer patients comprised an equally high proportion of men (94 vs 92%, respectively). The most common location of oropharyngeal Tis tumors was the posterior wall (32%), whereas T1-4 tumors were most commonly found on the lateral wall (36%). In hypopharyngeal cancer, both Tis and T1-4 were most commonly located in the pyriform sinus (62%). The proportion of Tis tumors diagnosed at individual institutions showed a positive correlation with the number of endoscopic treatments (r = 0.32, P squamous cell carcinoma patients in Japan. Further improvements in early diagnosis and standardized treatments are warranted.

  13. Superficial effects during the activation of zirconium AB2 alloys

    International Nuclear Information System (INIS)

    Zerbino, J; Visitin, A; Triaca, W

    2005-01-01

    The activation of zirconium nickel alloys with and without the addition of chromium and titanium is investigated through electrochemical and optical techniques.These alloys show high hydrogen absorption capacity and are extensively used in metal hydride batteries.Recent investigations in aqueous 1 M KOH indicate oxide layer growth and occlusion of hydrogen species in the alloys during the application of different cathodic potential programmes currently used in the activation process.In this research several techniques such as voltammetry, ellipsometry, energy dispersive analysis of X-rays EDAX, and scanning electron microscopy SEM are applied on the polished massive alloy Zr 1 -xTi x , x=0.36 y 0.43, and Zr 1 -xTi x CrNi, x=0.1,0.2 y 0.4.Data analysis shows that the stability, compactness and structure of the passive layers are strongly dependent on the applied potential programme.The alloy activation depends on the formation of deepen crevices that remain after a new polishing. Microscopic observation shows increase in the crevices thickness after the cathodic sweep potential cycling, which produces fragmentation of the grains and oxide growth during the activation process.This indicates metal breaking and intergranular dissolution that take place together with oxide and hydride formation.In some cases the resultant crevice thickness is one or two orders higher than that of the superficial oxide growth indicating intergranular localised corrosion

  14. Experimental Study of Phlebitis Ointment Administration in Acute Superficial Thrombophlebitis

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    Guangzong Li

    2018-01-01

    Full Text Available Acute superficial thrombophlebitis is a venous system disease. Animal models with mannitol induced phlebitis were treated with an orally administered “phlebitis ointment.” 24 rabbits were randomly divided into 4 groups. The therapy group was treated with “phlebitis ointment” and a control group received “Mai Luo Shu Tong granules.” Levels of blood TNF-α, IL-6, CRP, and IL-1β were measured. The tissue expression levels of NF-КBp65 and PKC genes were evaluated. The therapy group showed a better improvement of the clinical status and similar vascular morphology than the control group. A blank group showed no vascular changes through pathological investigation. In contrast, significant vascular changes were seen in the model group. The control group showed slight vascular modifications. Small thrombi could be found in the lumen despite the intact tunica intima. Both control and therapy group showed less inflammatory cells infiltration than the model group and upregulation of NF-КBp65 and PKC genes. The phlebitis ointment reduced the levels of necrosis factor-α, interleukin-6, C-reactive protein, and interleukin-1ß. The expressions of NF-КBp65 and PKC genes, which are the primary mechanisms underlying the development of thrombophlebitis, were improved significantly in tissues of both therapy group and control group.

  15. Superficial vessel reconstruction with a multiview camera system

    Science.gov (United States)

    Marreiros, Filipe M. M.; Rossitti, Sandro; Karlsson, Per M.; Wang, Chunliang; Gustafsson, Torbjörn; Carleberg, Per; Smedby, Örjan

    2016-01-01

    Abstract. We aim at reconstructing superficial vessels of the brain. Ultimately, they will serve to guide the deformation methods to compensate for the brain shift. A pipeline for three-dimensional (3-D) vessel reconstruction using three mono-complementary metal-oxide semiconductor cameras has been developed. Vessel centerlines are manually selected in the images. Using the properties of the Hessian matrix, the centerline points are assigned direction information. For correspondence matching, a combination of methods was used. The process starts with epipolar and spatial coherence constraints (geometrical constraints), followed by relaxation labeling and an iterative filtering where the 3-D points are compared to surfaces obtained using the thin-plate spline with decreasing relaxation parameter. Finally, the points are shifted to their local centroid position. Evaluation in virtual, phantom, and experimental images, including intraoperative data from patient experiments, shows that, with appropriate camera positions, the error estimates (root-mean square error and mean error) are ∼1  mm. PMID:26759814

  16. Internet: Más Allá de la superficie

    Directory of Open Access Journals (Sweden)

    Diego Andrés Tovar Cardozo

    2015-01-01

    Full Text Available El presente artículo expone un enfoque técnico de la Internet que da a conocer algunas características de su estructura, la cual muestra dos sectores distintos en donde se puede encontrar la información, es decir, esta se encuentra distribuida en niveles o capas de manera parcial o restringida, integrada o desagregada, todo ello dependiendo de lo que el usuario requiera realmente. La forma correcta para acceder a la información requerida está en saber a dónde ir a buscarla, la web profunda es información no visible para el usuario común que no tiene la habilidad de búsqueda, por lo cual se hace necesario verificar el tipo, la calidad y veracidad de la información a obtener antes de proceder a usar la información encontrada con los buscadores tradicionales. Acceder a Internet sin saber que existe una falsa cortina utilizada por la mayoría de los buscadores convencionales, es limitar las capacidades de navegar en un océano de información valiosa, en ello radica la importancia de conocer la web desde lo superficial (Surface Web a lo profundo (Deep Web.

  17. Inmersiones condicionadas de grafos en superficies y seudosuperficies.

    Directory of Open Access Journals (Sweden)

    Fedriani Martel, E. M.

    1999-01-01

    Full Text Available En este trabajo se procede a recapitular resultados conocidos sobre el problema de caracterizar los grafos que admiten inmersiones en superficies y en seudosuperficies con todos los vértices en la misma cara y se da una caracterización original de los grafos con dicha propiedad en seudosuperficies que surgen de manera natural y que han sido tratadas en la literatura especializada. Se comienza desarrollando algunos conceptos necesarios para la exposición de los problemas que se tratan, pero se suponen conocidos otros básicos de la Teoría de Grafos. Los resultados relacionados llevan de forma consecuente al planteamiento de otros problemas cuyas resoluciones originales también se incluyen. En cuanto a los objetivos finales que se pretenden lograr con esta comunicación, se basan, sobre todo, en aprovechar los resultados obtenidos por la Teoría de Grafos para resolver problemas pertenecientes a otras áreas, como la Economía. Se termina exponiendo un motivo topológico por el que estos estudios sobre las inmersiones y las inmersiones peri-S se hallan próximas a la Economía.

  18. Fine-needle aspiration cytology in children with superficial lymphadenopathy

    Directory of Open Access Journals (Sweden)

    F. De Corti

    2014-04-01

    Full Text Available Introduction: In pediatric population Fine-Needle Aspiration Citology (FNAC is slowly gaining acceptance in clinical management of Superficial Lymphadenopathy (SL. Our experience adds some data about the usefulness of this technique in diagnosing the cause of a SL and therefore guiding further treatment. Patients and Methods: 238 FNAC were performed in 217 patients with SL, observed at our Institution from 2002 to 2006. The neck was the most frequent localization. The results were available within few hours. In cases of granulomatous findings, the samples were processed for microbiological and PCR test, in order to identify Mycobacteria. Results: 174 were reactive lesions, 38 granulomatous lymphadenopathies, 24 malignant lesions, 2 specimens inadequate for diagnosis. Among the 174 reactive SL, 22 required an incisional biopsy after 1 month follow-up. Among the granulomatous lymphadenopathies, 13 children with Cat-Scratch Disease recovered, 25 with Mycobacteria infection underwent surgical excision. For 24 malignant lesions, the diagnosis was confirmed by further biopsy. Two false negative and no false positive were detected (sensitivity 92%, specificity 100%. No complications were encountered. Conclusions: In our experience FNAC, performed by experienced cytopathologist, has revealed to be a fast, safe, non invasive and inexpensive method to achieve diagnosis in persistent SL. The use of FNAC gave us the possibility to select patients for further investigation and/or surgical treatment. Incisional biopsy remains necessary to confirm the diagnosis in case of malignancy or doubtful lesions.

  19. Triterpenes for Well-Balanced Scar Formation in Superficial Wounds

    Directory of Open Access Journals (Sweden)

    Stefan Kindler

    2016-08-01

    Full Text Available Triterpenes are demonstrably effective for accelerating re-epithelialisation of wounds and known to improve scar formation for superficial lesions. Among the variety of triterpenes, betuline is of particular medical interest. Topical betuline gel (TBG received drug approval in 2016 from the European Commission as the first topical therapeutic agent with the proven clinical benefit of accelerating wound healing. Two self-conducted randomized intra-individual comparison clinical studies with a total of 220 patients involved in TBG treatment of skin graft surgical wounds have been screened for data concerning the aesthetic aspect of wound healing. Three months after surgery wound treatment with TBG resulted in about 30% of cases with more discreet scars, and standard of care in about 10%. Patients themselves appreciate the results of TBG after 3 months even more (about 50% compared to standard of care (about 10%. One year after surgery, the superiority of TBG counts for about 25% in comparison with about 10%, and from the patients’ point of view, for 25% compared to 4% under standard of care. In the majority of wound treatment cases, there is no difference visible between TBG treatment and standard of care after 1 year of scar formation. However, in comparison, TBG still offers a better chance for discreet scars and therefore happens to be superior in good care of wounds.

  20. Multivectored Superficial Muscular Aponeurotic System Suspension for Facial Paralysis.

    Science.gov (United States)

    Leach, Garrison; Kurnik, Nicole; Joganic, Jessica; Joganic, Edward

    2017-06-01

    Facial paralysis is a devastating condition that may cause severe cosmetic and functional deformities. In this study we describe our technique for superficial muscular aponeurotic system (SMAS) suspension using barbed suture and compare the vectors of suspension in relation to the underlying musculature. This study also quantifies the improvements in postoperative symmetry using traditional anthropologic landmarks. The efficacy of this procedure for improving facial paralysis was determined by comparing anthropometric indices and using Procrustes distance between 4 groupings of homologous landmarks plotted on each patient's preoperative and postoperative photos. Geometric morphometrics was used to evaluate change in facial shape and improvement in symmetry postoperatively.To analyze the vector of suspension in relation to the underlying musculature, specific anthropologic landmarks were used to calculate the vector of the musculature in 3 facial hemispheres from cadaveric controls against the vector of repair in our patients. Ten patients were included in our study. Subjectively, great improvement in functional status was achieved. Geometric morphometric analysis demonstrated a statistically significant improvement in facial symmetry. Cadaveric dissection demonstrated that the suture should be placed in the SMAS in vectors parallel to the underlying musculature to achieve these results. There were no complications in our study to date. In conclusion, multivectored SMAS suture suspension is an effective method for restoring static suspension of the face after facial paralysis. This method has the benefit of producing quick, reliable results with improved function, low cost, and low morbidity.

  1. DRENAGEM SUPERFICIAL DE SOLOS ARGILOSOS DO NORTE DO PARANÁ

    Directory of Open Access Journals (Sweden)

    Rogério Teixeira de Faria

    1999-08-01

    Full Text Available RESUMO Foi avaliado o desempenho de práticas de drenagem superficial e cobertura morta em uma área de várzea do norte do Paraná. Antes e após o cultivo do arroz, foram comparadas as taxas de drenagem em tratamentos com e sem drenos torpedo e determinadas a vazão e a obstrução dos condutos. Sob o cultivo do feijoeiro, foram avaliados os efeitos dos drenos torpedo e de "camas" na drenagem e de cobertura morta (palha de arroz na retenção de água do solo. Os resultados indicaram que os drenos torpedo foram mais eficientes se construídos com solo menos úmido, porém sua longevidade foi menor que um ano. Sob o feijoeiro, os drenos torpedo e cobertura morta mantiveram o lençol freático mais profundo e o solo com menor tensão de umidade, mas esses efeitos ocorreram abaixo da zona radicular (15 cm, resultando em baixa produtividade.

  2. Recurrence of superficial vein thrombosis in patients with varicose veins.

    Science.gov (United States)

    Karathanos, Christos; Spanos, Konstantinos; Saleptsis, Vassileios; Tsezou, Aspasia; Kyriakou, Despina; Giannoukas, Athanasios D

    2016-08-01

    To investigate which factors other than history of superficial vein thrombosis (SVT) are associated with recurrent spontaneous SVT episodes in patients with varicose veins (VVs). Patients with a history of spontaneous SVT and VVs were followed up for a mean period of 55 months. Demographics, comorbidities, and thrombophilia screening test were analyzed. Patients were grouped according to the clinical-etiology-anatomy-pathophysiology classification. A multiple logistic regression analysis with the forward likelihood ratio method was undertaken. Thirteen patients out of 97 had a recurrence SVT episode during the follow-up period. All those patients were identified to have a thrombophilia defect. Protein C and S, antithrombin, and plasminogen deficiencies were more frequently present in patients without recurrence. Gene mutations were present in 38% in the nonrecurrence group and 77% in the recurrence group. After logistic regression analysis, patients with dislipidemia and mutation in prothrombin G20210A (FII) had an increased risk for recurrence by 5.4-fold and 4.6-fold, respectively. No deep vein thrombosis or pulmonary embolism occurred. Dislipidemia and gene mutations of F II are associated with SVT recurrence in patients with VVs. A selection of patients may benefit from anticoagulation in the short term and from VVs intervention in the long term. © The Author(s) 2015.

  3. Comparisons of actin filament disruptors and Rho kinase inhibitors as potential antiglaucoma medications

    OpenAIRE

    Tian, Baohe; Kaufman, Paul L

    2012-01-01

    Dynamics of the actin cytoskeleton in the trabecular meshwork play a crucial role in the regulation of trabecular outflow resistance. The actin filament disruptors and Rho kinase inhibitors affect the dynamics of the actomyosin system by either disrupting the actin filaments or inhibiting the Rho kinase-activated cellular contractility. Both approaches induce similar morphological changes and resistance decreases in the trabecular outflow pathway, and thus both have potential as antiglaucoma ...

  4. Cell stress promotes the association of phosphorylated HspB1 with F-actin.

    Directory of Open Access Journals (Sweden)

    Joseph P Clarke

    Full Text Available Previous studies have suggested that the small heat shock protein, HspB1, has a direct influence on the dynamics of cytoskeletal elements, in particular, filamentous actin (F-actin polymerization. In this study we have assessed the influence of HspB1 phosphorylation on its interaction(s with F-actin. We first determined the distribution of endogenous non-phosphorylated HspB1, phosphorylated HspB1 and F-actin in neuroendocrine PC12 cells by immunocytochemistry and confocal microscopy. We then investigated a potential direct interaction between HspB1 with F-actin by precipitating F-actin directly with biotinylated phalloidin followed by Western analyses; the reverse immunoprecipitation of HspB1 was also carried out. The phosphorylation influence of HspB1 in this interaction was investigated by using pharmacologic inhibition of p38 MAPK. In control cells, HspB1 interacts with F-actin as a predominantly non-phosphorylated protein, but subsequent to stress there is a redistribution of HspB1 to the cytoskeletal fraction and a significantly increased association of pHspB1 with F-actin. Our data demonstrate HspB1 is found in a complex with F-actin both in phosphorylated and non-phosphorylated forms, with an increased association of pHspB1 with F-actin after heat stress. Overall, our study combines both cellular and biochemical approaches to show cellular localization and direct demonstration of an interaction between endogenous HspB1 and F-actin using methodolgy that specifically isolates F-actin.

  5. G-actin sequestering protein thymosin-β4 regulates the activity of myocardin-related transcription factor.

    Science.gov (United States)

    Morita, Tsuyoshi; Hayashi, Ken'ichiro

    2013-08-02

    Myocardin-related transcription factors (MRTFs) are robust coactivators of serum response factor (SRF). MRTFs contain three copies of the RPEL motif at their N-terminus, and they bind to monomeric globular actin (G-actin). Previous studies illustrate that G-actin binding inhibits MRTF activity by preventing the MRTFs nuclear accumulation. In the living cells, the majority of G-actin is sequestered by G-actin binding proteins that prevent spontaneous actin polymerization. Here, we demonstrate that the most abundant G-actin sequestering protein thymosin-β4 (Tβ4) was involved in the regulation of subcellular localization and activity of MRTF-A. Tβ4 competed with MRTF-A for G-actin binding; thus, interfering with G-actin-MRTF-A complex formation. Tβ4 overexpression induced the MRTF-A nuclear accumulation and activation of MRTF-SRF signaling. The activation rate of MRTF-A by the Tβ4 mutant L17A, whose affinity for G-actin is very low, was lower than that by wild-type Tβ4. In contrast, the β-actin mutant 3DA, which has a lower affinity for Tβ4, more effectively suppressed MRTF-A activity than wild-type β-actin. Furthermore, ectopic Tβ4 increased the endogenous expression of SRF-dependent actin cytoskeletal genes. Thus, Tβ4 is an important MRTF regulator that controls the G-actin-MRTFs interaction. Copyright © 2013 Elsevier Inc. All rights reserved.

  6. Actin-myosin network is required for proper assembly of influenza virus particles

    Energy Technology Data Exchange (ETDEWEB)

    Kumakura, Michiko; Kawaguchi, Atsushi, E-mail: ats-kawaguchi@md.tsukuba.ac.jp; Nagata, Kyosuke, E-mail: knagata@md.tsukuba.ac.jp

    2015-02-15

    Actin filaments are known to play a central role in cellular dynamics. After polymerization of actin, various actin-crosslinking proteins including non-muscle myosin II facilitate the formation of spatially organized actin filament networks. The actin-myosin network is highly expanded beneath plasma membrane. The genome of influenza virus (vRNA) replicates in the cell nucleus. Then, newly synthesized vRNAs are nuclear-exported to the cytoplasm as ribonucleoprotein complexes (vRNPs), followed by transport to the beneath plasma membrane where virus particles assemble. Here, we found that, by inhibiting actin-myosin network formation, the virus titer tends to be reduced and HA viral spike protein is aggregated on the plasma membrane. These results indicate that the actin-myosin network plays an important role in the virus formation. - Highlights: • Actin-myosin network is important for the influenza virus production. • HA forms aggregations at the plasma membrane in the presence of blebbistatin. • M1 is recruited to the budding site through the actin-myosin network.

  7. Profilin-Dependent Nucleation and Assembly of Actin Filaments Controls Cell Elongation in Arabidopsis1[OPEN

    Science.gov (United States)

    Cao, Lingyan; Blanchoin, Laurent; Staiger, Christopher J.

    2016-01-01

    Actin filaments in plant cells are incredibly dynamic; they undergo incessant remodeling and assembly or disassembly within seconds. These dynamic events are choreographed by a plethora of actin-binding proteins, but the exact mechanisms are poorly understood. Here, we dissect the contribution of Arabidopsis (Arabidopsis thaliana) PROFILIN1 (PRF1), a conserved actin monomer-binding protein, to actin organization and single filament dynamics during axial cell expansion of living epidermal cells. We found that reduced PRF1 levels enhanced cell and organ growth. Surprisingly, we observed that the overall frequency of nucleation events in prf1 mutants was dramatically decreased and that a subpopulation of actin filaments that assemble at high rates was reduced. To test whether profilin cooperates with plant formin proteins to execute actin nucleation and rapid filament elongation in cells, we used a pharmacological approach. Here, we used Small Molecule Inhibitor of Formin FH2 (SMIFH2), after validating its mode of action on a plant formin in vitro, and observed a reduced nucleation frequency of actin filaments in live cells. Treatment of wild-type epidermal cells with SMIFH2 mimicked the phenotype of prf1 mutants, and the nucleation frequency in prf1-2 mutant was completely insensitive to these treatments. Our data provide compelling evidence that PRF1 coordinates the stochastic dynamic properties of actin filaments by modulating formin-mediated actin nucleation and assembly during plant cell expansion. PMID:26574597

  8. Actin-myosin network is required for proper assembly of influenza virus particles

    International Nuclear Information System (INIS)

    Kumakura, Michiko; Kawaguchi, Atsushi; Nagata, Kyosuke

    2015-01-01

    Actin filaments are known to play a central role in cellular dynamics. After polymerization of actin, various actin-crosslinking proteins including non-muscle myosin II facilitate the formation of spatially organized actin filament networks. The actin-myosin network is highly expanded beneath plasma membrane. The genome of influenza virus (vRNA) replicates in the cell nucleus. Then, newly synthesized vRNAs are nuclear-exported to the cytoplasm as ribonucleoprotein complexes (vRNPs), followed by transport to the beneath plasma membrane where virus particles assemble. Here, we found that, by inhibiting actin-myosin network formation, the virus titer tends to be reduced and HA viral spike protein is aggregated on the plasma membrane. These results indicate that the actin-myosin network plays an important role in the virus formation. - Highlights: • Actin-myosin network is important for the influenza virus production. • HA forms aggregations at the plasma membrane in the presence of blebbistatin. • M1 is recruited to the budding site through the actin-myosin network

  9. Cooperative and non-cooperative conformational changes of F-actin induced by cofilin

    Energy Technology Data Exchange (ETDEWEB)

    Aihara, Tomoki; Oda, Toshiro, E-mail: toda@spring8.or.jp

    2013-05-31

    Highlights: •Mobility of MTSL attached to C374 in F-actin became high upon addition of cofilin. •Change of motility of MTSL attached to C374 with cofilin-binding was cooperative. •Mobility of MTSL attached to V43C in F-actin became high upon addition of cofilin. •Change of motility of MTSL attached to V43C with cofilin-binding was linear. -- Abstract: Cofilin is an actin-binding protein that promotes F-actin depolymerization. It is well-known that cofilin-coated F-actin is more twisted than naked F-actin, and that the protomer is more tilted. However, the means by which the local changes induced by the binding of individual cofilin proteins proceed to the global conformational changes of the whole F-actin molecule remain unknown. Here we investigated the cofilin-induced changes in several parts of F-actin, through site-directed spin-label electron paramagnetic resonance spectroscopy analyses of recombinant actins containing single reactive cysteines. We found that the global, cooperative conformational changes induced by cofilin-binding, which were detected by the spin-label attached to the Cys374 residue, occurred without the detachment of the D-loop in subdomain 2 from the neighboring protomer. The two processes of local and global changes do not necessarily proceed in sequence.

  10. Organization and dynamics of the actin cytoskeleton during dendritic spine morphological remodeling.

    Science.gov (United States)

    Chazeau, Anaël; Giannone, Grégory

    2016-08-01

    In the central nervous system, most excitatory post-synapses are small subcellular structures called dendritic spines. Their structure and morphological remodeling are tightly coupled to changes in synaptic transmission. The F-actin cytoskeleton is the main driving force of dendritic spine remodeling and sustains synaptic plasticity. It is therefore essential to understand how changes in synaptic transmission can regulate the organization and dynamics of actin binding proteins (ABPs). In this review, we will provide a detailed description of the organization and dynamics of F-actin and ABPs in dendritic spines and will discuss the current models explaining how the actin cytoskeleton sustains both structural and functional synaptic plasticity.

  11. Investigating sub-spine actin dynamics in rat hippocampal neurons with super-resolution optical imaging.

    Directory of Open Access Journals (Sweden)

    Vedakumar Tatavarty

    Full Text Available Morphological changes in dendritic spines represent an important mechanism for synaptic plasticity which is postulated to underlie the vital cognitive phenomena of learning and memory. These morphological changes are driven by the dynamic actin cytoskeleton that is present in dendritic spines. The study of actin dynamics in these spines traditionally has been hindered by the small size of the spine. In this study, we utilize a photo-activation localization microscopy (PALM-based single-molecule tracking technique to analyze F-actin movements with approximately 30-nm resolution in cultured hippocampal neurons. We were able to observe the kinematic (physical motion of actin filaments, i.e., retrograde flow and kinetic (F-actin turn-over dynamics of F-actin at the single-filament level in dendritic spines. We found that F-actin in dendritic spines exhibits highly heterogeneous kinematic dynamics at the individual filament level, with simultaneous actin flows in both retrograde and anterograde directions. At the ensemble level, movements of filaments integrate into a net retrograde flow of approximately 138 nm/min. These results suggest a weakly polarized F-actin network that consists of mostly short filaments in dendritic spines.

  12. Investigating sub-spine actin dynamics in rat hippocampal neurons with super-resolution optical imaging.

    Science.gov (United States)

    Tatavarty, Vedakumar; Kim, Eun-Ji; Rodionov, Vladimir; Yu, Ji

    2009-11-09

    Morphological changes in dendritic spines represent an important mechanism for synaptic plasticity which is postulated to underlie the vital cognitive phenomena of learning and memory. These morphological changes are driven by the dynamic actin cytoskeleton that is present in dendritic spines. The study of actin dynamics in these spines traditionally has been hindered by the small size of the spine. In this study, we utilize a photo-activation localization microscopy (PALM)-based single-molecule tracking technique to analyze F-actin movements with approximately 30-nm resolution in cultured hippocampal neurons. We were able to observe the kinematic (physical motion of actin filaments, i.e., retrograde flow) and kinetic (F-actin turn-over) dynamics of F-actin at the single-filament level in dendritic spines. We found that F-actin in dendritic spines exhibits highly heterogeneous kinematic dynamics at the individual filament level, with simultaneous actin flows in both retrograde and anterograde directions. At the ensemble level, movements of filaments integrate into a net retrograde flow of approximately 138 nm/min. These results suggest a weakly polarized F-actin network that consists of mostly short filaments in dendritic spines.

  13. Dual effect of pseudorabies virus growth factor (PRGF) displayed on actin cytoskeleton.

    Science.gov (United States)

    Urbancíková, M; Vozárová, G; Lesko, J; Golais, F

    1999-10-01

    Pseudorabies virus growth factor (PRGF) was shown to possess transforming activity as well as transformation repressing activity in in vitro systems. In order to better understand these phenomena we studied actin cytoskeleton and its alterations induced by PRGF using normal human fibroblasts VH-10 and transformed cell line HeLa. For specific detection of filamentous actin cells were stained with phalloidin conjugated with fluorescein isothiocyanate (FITC)-phalloidin. PRGF was applied to VH-10 cells for various length of time from 10 min up to 48 h. The effect was very fast and changes in actin filament composition could be detected already after 10 min. In comparison to untreated cells the staining of treated cells was more diffuse and a number of actin microfilaments in individual stress fibers became reduced. After 30 min thick short actin bundles appeared in the perinuclear region. A 24-h exposure resulted in a large reduction of actin bundles. After additional 24 h a partial restoration of actin cytoskeleton in cells was observed. In transformed HeLa cells PRGF induced opposite process than in normal cells: the number of filamentous actin structures increased. We hypothesise that PRGF may act as a transcription-like factor and may initiate changes in gene expression which consequently result in actin cytoskeleton alterations.

  14. TWISTED DWARF1 Mediates the Action of Auxin Transport Inhibitors on Actin Cytoskeleton Dynamics

    Science.gov (United States)

    Bailly, Aurelien; Zwiewka, Marta; Sovero, Valpuri; Ge, Pei; Aryal, Bibek; Hao, Pengchao; Linnert, Miriam; Burgardt, Noelia Inés; Lücke, Christian; Weiwad, Matthias; Michel, Max; Weiergräber, Oliver H.; Pollmann, Stephan; Azzarello, Elisa; Fukao, Yoichiro; Hoffmann, Céline; Wedlich-Söldner, Roland

    2016-01-01

    Plant growth and architecture is regulated by the polar distribution of the hormone auxin. Polarity and flexibility of this process is provided by constant cycling of auxin transporter vesicles along actin filaments, coordinated by a positive auxin-actin feedback loop. Both polar auxin transport and vesicle cycling are inhibited by synthetic auxin transport inhibitors, such as 1-N-naphthylphthalamic acid (NPA), counteracting the effect of auxin; however, underlying targets and mechanisms are unclear. Using NMR, we map the NPA binding surface on the Arabidopsis thaliana ABCB chaperone TWISTED DWARF1 (TWD1). We identify ACTIN7 as a relevant, although likely indirect, TWD1 interactor, and show TWD1-dependent regulation of actin filament organization and dynamics and that TWD1 is required for NPA-mediated actin cytoskeleton remodeling. The TWD1-ACTIN7 axis controls plasma membrane presence of efflux transporters, and as a consequence act7 and twd1 share developmental and physiological phenotypes indicative of defects in auxin transport. These can be phenocopied by NPA treatment or by chemical actin (de)stabilization. We provide evidence that TWD1 determines downstream locations of auxin efflux transporters by adjusting actin filament debundling and dynamizing processes and mediating NPA action on the latter. This function appears to be evolutionary conserved since TWD1 expression in budding yeast alters actin polarization and cell polarity and provides NPA sensitivity. PMID:27053424

  15. Plant vegetative and animal cytoplasmic actins share functional competence for spatial development with protists.

    Science.gov (United States)

    Kandasamy, Muthugapatti K; McKinney, Elizabeth C; Roy, Eileen; Meagher, Richard B

    2012-05-01

    Actin is an essential multifunctional protein encoded by two distinct ancient classes of genes in animals (cytoplasmic and muscle) and plants (vegetative and reproductive). The prevailing view is that each class of actin variants is functionally distinct. However, we propose that the vegetative plant and cytoplasmic animal variants have conserved functional competence for spatial development inherited from an ancestral protist actin sequence. To test this idea, we ectopically expressed animal and protist actins in Arabidopsis thaliana double vegetative actin mutants that are dramatically altered in cell and organ morphologies. We found that expression of cytoplasmic actins from humans and even a highly divergent invertebrate Ciona intestinalis qualitatively and quantitatively suppressed the root cell polarity and organ defects of act8 act7 mutants and moderately suppressed the root-hairless phenotype of act2 act8 mutants. By contrast, human muscle actins were unable to support prominently any aspect of plant development. Furthermore, actins from three protists representing Choanozoa, Archamoeba, and green algae efficiently suppressed all the phenotypes of both the plant mutants. Remarkably, these data imply that actin's competence to carry out a complex suite of processes essential for multicellular development was already fully developed in single-celled protists and evolved nonprogressively from protists to plants and animals.

  16. A function for filamentous alpha-smooth muscle actin: Retardation of motility in human breast fibroblasts

    DEFF Research Database (Denmark)

    Rønnov-Jessen, Lone; Petersen, Ole William

    1996-01-01

    .8 and 3.0 microns/h, respectively. To knock out the alpha-sm actin protein, several antisense phosphorothioate oligodeoxynucleotide (ODNs) were tested. One of these, 3'UTI, which is complementary to a highly evolutionary conserved 3' untranslated (3'UT) sequence of alpha-sm actin mRNA, was found to block...... alpha-sm actin synthesis completely without affecting the synthesis of any other proteins as analyzed by two-dimensional gel electrophoresis. Targeting by antisense 3'UTI significantly increased motility compared with the corresponding sense ODN. alpha-Sm actin inhibition also led to the formation...

  17. Plant villin, lily P-135-ABP, possesses G-actin binding activity and accelerates the polymerization and depolymerization of actin in a Ca2+-sensitive manner.

    Science.gov (United States)

    Yokota, Etsuo; Tominaga, Motoki; Mabuchi, Issei; Tsuji, Yasunori; Staiger, Christopher J; Oiwa, Kazuhiro; Shimmen, Teruo

    2005-10-01

    From germinating pollen of lily, two types of villins, P-115-ABP and P-135-ABP, have been identified biochemically. Ca(2+)-CaM-dependent actin-filament binding and bundling activities have been demonstrated for both villins previously. Here, we examined the effects of lily villins on the polymerization and depolymerization of actin. P-115-ABP and P-135-ABP present in a crude protein extract prepared from germinating pollen bound to a DNase I affinity column in a Ca(2+)-dependent manner. Purified P-135-ABP reduced the lag period that precedes actin filament polymerization from monomers in the presence of either Ca(2+) or Ca(2+)-CaM. These results indicated that P-135-ABP can form a complex with G-actin in the presence of Ca(2+) and this complex acts as a nucleus for polymerization of actin filaments. However, the nucleation activity of P-135-ABP is probably not relevant in vivo because the assembly of G-actin saturated with profilin, a situation that mimics conditions found in pollen, was not accelerated in the presence of P-135-ABP. P-135-ABP also enhanced the depolymerization of actin filaments during dilution-mediated disassembly. Growth from filament barbed ends in the presence of Ca(2+)-CaM was also prevented, consistent with filament capping activity. These results suggested that lily villin is involved not only in the arrangement of actin filaments into bundles in the basal and shank region of the pollen tube, but also in regulating and modulating actin dynamics through its capping and depolymerization (or fragmentation) activities in the apical region of the pollen tube, where there is a relatively high concentration of Ca(2+).

  18. External stimulation strength controls actin response dynamics in Dictyostelium cells

    Science.gov (United States)

    Hsu, Hsin-Fang; Westendorf, Christian; Tarantola, Marco; Zykov, Vladimir; Bodenschatz, Eberhard; Beta, Carsten

    2015-03-01

    Self-sustained oscillation and the resonance frequency of the cytoskeletal actin polymerization/depolymerization have recently been observed in Dictyostelium, a model system for studying chemotaxis. Here we report that the resonance frequency is not constant but rather varies with the strength of external stimuli. To understand the underlying mechanism, we analyzed the polymerization and depolymerization time at different levels of external stimulation. We found that polymerization time is independent of external stimuli but the depolymerization time is prolonged as the stimulation increases. These observations can be successfully reproduced in the frame work of our time delayed differential equation model.

  19. Analyses of superficial and depth doses in intraoral radiology

    Energy Technology Data Exchange (ETDEWEB)

    Silva Santos de Oliveira, C.; Morais, R.P. de; Nascimento Souza, D. do [Universidade Federal de Sergipe - CCET - Dept. de Fisica, Sao Cristovao, SE (Brazil)

    2006-07-01

    In this work dosimetric analysis using thermoluminescence technique to study the beams characteristics of x-rays employed in dental radiology has been carried out. The obtained results with CaSO{sub 4}:Dy thermoluminescent dosimeters (TLD) were compared to the doses obtained with parallel-plates ionization chamber. Dosimetric evaluations were also done using radiographic films of large dimensions. The x-rays equipments analyzed were installed in the radiological services of Odontology Department of Sergipe Federal University (U.F.S.). Depending on the anatomical region to be examined the proper exposure time was select, for a fix voltage of 70 kV. The results with TLD and ionization chamber have been determined to female and male individuals. The intraoral regions analysed were the peri apical of the incisors, molar and pre-molar teeth and the occlusive region. These regions were simulated using acrylic plates absorbers installed on the film packet holder. The evaluation of the depth doses in the intraoral tissue was obtained using different acrylic plate thickness. The air kerma values have been evaluated with the ionization chamber located in the dental cone exit of the x-rays equipments. The integrated areas of the thermoluminescent glow curves showed coherent values when compared to the ones obtained with the ionization chamber and both methods presented a linear dependence with the exposition time. The analyses with films have allowed the evaluation of the beam scattering in the simulator apparatus. The studies had proven that the analysis of superficial dose and in depth used in dental radiology can be carried with thermoluminescent dosimeters. (authors)

  20. Teledetection applied to the superficial oceanography phenomena study

    International Nuclear Information System (INIS)

    Melo, Jeimmy

    2002-01-01

    The present document is consider the physical and biological variables important part in the integration of the climatic system, like improvement to the knowledge of the present marine processes in the Colombian marine areas by means of the advantage of the technological resources available at the present time for the obtaining of information in real time of physical and biological processes, agreed to the constitutional mandate of sustainable development. The images of color of the ocean provided by located remote sensors in the satellites allow considering primary the organic productivity of a synoptic way, in great areas. This is of great importance to more effectively support the rational handling of the fishing resources and to make a contribution on the paper of the ocean in the global climatic change. The marine phytoplankton can significantly alter to the carbon interchange in interactions ocean-atmosphere. The marine photosynthesis reduces the partial pressure of superficial water CO 2 , with the result of which a portion of organic carbon just formed (new productivity) sinks outside the afotica zone. The previous thing induces a net effect of carbon flow from the atmosphere to the ocean. One of the potential applications of the use of the images of satellite of color of the ocean is the calculation of the chlorophyll concentration and its relation with the sobreexperience of larvae of fish of commercial interest. Different types from developed mathematical models for concerning oceanic waters in the accomplishment of estimations of primary productivity with base in color images of the ocean exist mainly - time out - for that reason, the necessity of develop model similar for the calculation of the productivity primary of the phytoplankton of the Colombian marine area

  1. Analyses of superficial and depth doses in intraoral radiology

    International Nuclear Information System (INIS)

    Silva Santos de Oliveira, C.; Morais, R.P. de; Nascimento Souza, D. do

    2006-01-01

    In this work dosimetric analysis using thermoluminescence technique to study the beams characteristics of x-rays employed in dental radiology has been carried out. The obtained results with CaSO 4 :Dy thermoluminescent dosimeters (TLD) were compared to the doses obtained with parallel-plates ionization chamber. Dosimetric evaluations were also done using radiographic films of large dimensions. The x-rays equipments analyzed were installed in the radiological services of Odontology Department of Sergipe Federal University (U.F.S.). Depending on the anatomical region to be examined the proper exposure time was select, for a fix voltage of 70 kV. The results with TLD and ionization chamber have been determined to female and male individuals. The intraoral regions analysed were the peri apical of the incisors, molar and pre-molar teeth and the occlusive region. These regions were simulated using acrylic plates absorbers installed on the film packet holder. The evaluation of the depth doses in the intraoral tissue was obtained using different acrylic plate thickness. The air kerma values have been evaluated with the ionization chamber located in the dental cone exit of the x-rays equipments. The integrated areas of the thermoluminescent glow curves showed coherent values when compared to the ones obtained with the ionization chamber and both methods presented a linear dependence with the exposition time. The analyses with films have allowed the evaluation of the beam scattering in the simulator apparatus. The studies had proven that the analysis of superficial dose and in depth used in dental radiology can be carried with thermoluminescent dosimeters. (authors)

  2. Terbinafine-loaded wound dressing for chronic superficial fungal infections.

    Science.gov (United States)

    Paskiabi, Farnoush Asghari; Bonakdar, Shahin; Shokrgozar, Mohammad Ali; Imani, Mohammad; Jahanshiri, Zahra; Shams-Ghahfarokhi, Masoomeh; Razzaghi-Abyaneh, Mehdi

    2017-04-01

    In spite of developing new drugs and modern formulations, the treatments of chronic fungal infections are still challenging. Fibrous wound dressings are new suggestions for the treatment of chronic superficial infections. In the present study, we formulated an antifungal agent, terbinafine hydrochloride (TFH), which is a hydrophobic drug, in wound dressings prepared by electrospun polycaprolactone, polycaprolactone/gelatin (50:50 w/w) and gelatin. To obtain more water-stable meshes, the preparations were treated by glutaraldehyde and their properties were determined before and after treatment. The morphology of fibrous meshes was observed by scanning electron microscopy. Drug loading efficiency and release rate were measured by high performance liquid chromatography (HPLC) and the release rate was monitored for 144h. Antifungal tests were performed on Trichophyton mentagrophytes, Aspergillus fumigatus and Candida albicans cultured on Muller-Hinton agar. The toxicity of the meshes was measured after 24h and 14days by MTT assay. Terbinafine loading of polycaprolactone/gelatin (50:50) was 100% and it released the highest amount of TFH too. In antifungal tests, all samples were able to hinderT. mentagrophytes and A. fumigatus but not C. albicans growth among them, polycaprolactone fibers made the largest inhibition zone. In MTT assay, none of prepared samples showed toxicity against L929 cells. Teken together, the prepared TFH-loaded PCL/gelatin electrospun meshes were able to release TFH slowly and in a steady state in time. With respect to no obvious cytotoxicity in MTT assay and stong antifungal activity toward T. mentagrophytesin vitro, these TFH-based meshes could be considered as potential candidates in clinical application as wound dressing for treatment of chronic dermatophytosis. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. HDR brachytherapy for superficial non-melanoma skin cancers

    International Nuclear Information System (INIS)

    Gauden, Ruth; Pracy, Martin; Avery, Anne-Marie; Hodgetts, Ian; Gauden, Stan

    2013-01-01

    Our initial experience using recommended high dose per fraction skin brachytherapy (BT) treatment schedules, resulted in poor cosmesis. This study aimed to assess in a prospective group of patients the use of Leipzig surface applicators for High Dose Rate (HDR) brachytherapy, for the treatment of small non-melanoma skin cancers (NMSC) using a protracted treatment schedule. Treatment was delivered by HDR brachytherapy with Leipzig applicators. 36Gy, prescribed to between 3 to 4mm, was given in daily 3Gy fractions. Acute skin toxicity was evaluated weekly during irradiation using the Radiation Therapy Oncology Group criteria. Local response, late skin effects and cosmetic results were monitored at periodic intervals after treatment completion. From March 2002, 200 patients with 236 lesions were treated. Median follow-up was 66 months (range 25–121 months). A total of 162 lesions were macroscopic, while in 74 cases, BT was given after resection because of positive microscopic margins. There were 121 lesions that were basal cell carcinomas, and 115 were squamous cell carcinomas. Lesions were located on the head and neck (198), the extremities (26) and trunk (12). Local control was 232/236 (98%). Four patients required further surgery to treat recurrence. Grade 1 acute skin toxicity was detected in 168 treated lesions (71%) and grade 2 in 81 (34%). Cosmesis was good or excellent in 208 cases (88%). Late skin hypopigmentation changes were observed in 13 cases (5.5%). Delivering 36Gy over 2 weeks to superficial NMSC using HDR brachytherapy is well tolerated and provides a high local control rate without significant toxicity.

  4. Anatomical and Electrophysiological Clustering of Superficial Medial Entorhinal Cortex Interneurons

    Science.gov (United States)

    2017-01-01

    Abstract Local GABAergic interneurons regulate the activity of spatially-modulated principal cells in the medial entorhinal cortex (MEC), mediating stellate-to-stellate connectivity and possibly enabling grid formation via recurrent inhibitory circuitry. Despite the important role interneurons seem to play in the MEC cortical circuit, the combination of low cell counts and functional diversity has made systematic electrophysiological studies of these neurons difficult. For these reasons, there remains a paucity of knowledge on the electrophysiological profiles of superficial MEC interneuron populations. Taking advantage of glutamic acid decarboxylase 2 (GAD2)-IRES-tdTomato and PV-tdTomato transgenic mice, we targeted GABAergic interneurons for whole-cell patch-clamp recordings and characterized their passive membrane features, basic input/output properties and action potential (AP) shape. These electrophysiologically characterized cells were then anatomically reconstructed, with emphasis on axonal projections and pial depth. K-means clustering of interneuron anatomical and electrophysiological data optimally classified a population of 106 interneurons into four distinct clusters. The first cluster is comprised of layer 2- and 3-projecting, slow-firing interneurons. The second cluster is comprised largely of PV+ fast-firing interneurons that project mainly to layers 2 and 3. The third cluster contains layer 1- and 2-projecting interneurons, and the fourth cluster is made up of layer 1-projecting horizontal interneurons. These results, among others, will provide greater understanding of the electrophysiological characteristics of MEC interneurons, help guide future in vivo studies, and may aid in uncovering the mechanism of grid field formation. PMID:29085901

  5. Use your eyes - metallographic versus superficial traces of the crafting of Bronze Age ornaments

    DEFF Research Database (Denmark)

    Nørgaard, Heide Wrobel

    and superficial archaeological examinations can be used to understand prehistoric craft. Furthermore, it will show that with an intensive knowledge about previous scientific investigations the superficial traces preserved can reveal a great amount of information and could talk for themselves. Taking as an example...... the early and middle Bronze Age bronze ornaments from North Germany, superficial traces of the metal crafting process, visible with the naked eye, will be compared with metallographic images. The aim of this presentation is to give an overview over the crafting traces everybody could see (with a bit...

  6. Unexpected diagnosis of superficial neurofibroma in a lesion with imaging features of a vascular malformation

    Energy Technology Data Exchange (ETDEWEB)

    O' Keefe, Patrick; Reid, Janet; Morrison, Stuart [Cleveland Clinic Foundation, Department of Radiology, Cleveland, OH (United States); Vidimos, Allison [Cleveland Clinic Foundation, Department of Dermatology, Cleveland, OH (United States); DiFiore, John [Cleveland Clinic Foundation, Department of Pediatric Surgery, Cleveland, OH (United States)

    2005-12-01

    Plexiform neurofibroma is a pathognomonic, often disabling feature of neurofibromatosis type I. Although the target-like appearance of deep plexiform neurofibroma on T2-weighted MRI has been well-described, a second superficial form of plexiform neurofibroma has differing imaging features. We report a 15-year-old boy who presented with multiple cutaneous lesions exhibiting clinical and imaging characteristics of a venolymphatic malformation. These lesions were histologically proved to represent superficial plexiform neurofibromas. We wish to emphasize the unique MR findings of superficial plexiform neurofibromas; these findings are different from the imaging characteristics of the deep form and can be confused with a low-flow vascular malformation. (orig.)

  7. Comportamiento a fatiga del acero sae 4140 usando alta rugosidad superficial y ambiente corrosivo

    OpenAIRE

    CEBALLOS, WILSON FERNANDO; GÓMEZ, ADOLFO LEÓN; CORONADO, JOHN JAIRO

    2010-01-01

    En este artículo se presenta el estudio del comportamiento a fatiga del acero SAE 4140 en condiciones diferentes: superficie pulida (acabado espejo), ambiente corrosivo (jugo de caña de azúcar), alta rugosidad superficial (superficie equivalente a la encontrada típicamente en los ejes de molino de caña) y una condición que involucra la combinación de las dos últimas. Las anteriores condiciones están presentes en los ejes de maza de molino de caña de azúcar durante el proceso de molienda. Se r...

  8. COMPORTAMIENTO A FATIGA DEL ACERO SAE 4140 USANDO ALTA RUGOSIDAD SUPERFICIAL Y AMBIENTE CORROSIVO

    OpenAIRE

    CEBALLOS, WILSON FERNANDO; GÓMEZ, ADOLFO LEÓN; CORONADO, JOHN JAIRO

    2010-01-01

    En este artículo se presenta el estudio del comportamiento a fatiga del acero SAE 4140 en condiciones diferentes: superficie pulida (acabado espejo), ambiente corrosivo (jugo de caña de azúcar), alta rugosidad superficial (superficie equivalente a la encontrada típicamente en los ejes de molino de caña) y una condición que involucra la combinación de las dos últimas. Las anteriores condiciones están presentes en los ejes de maza de molino de caña de azúcar durante el proceso de molienda. Se r...

  9. Actin cytoskeleton of chemotactic amoebae operates close to the onset of oscillations

    Science.gov (United States)

    Westendorf, Christian; Negrete, Jose; Bae, Albert J.; Sandmann, Rabea; Bodenschatz, Eberhard; Beta, Carsten

    2013-01-01

    The rapid reorganization of the actin cytoskeleton in response to external stimuli is an essential property of many motile eukaryotic cells. Here, we report evidence that the actin machinery of chemotactic Dictyostelium cells operates close to an oscillatory instability. When averaging the actin response of many cells to a short pulse of the chemoattractant cAMP, we observed a transient accumulation of cortical actin reminiscent of a damped oscillation. At the single-cell level, however, the response dynamics ranged from short, strongly damped responses to slowly decaying, weakly damped oscillations. Furthermore, in a small subpopulation, we observed self-sustained oscillations in the cortical F-actin concentration. To substantiate that an oscillatory mechanism governs the actin dynamics in these cells, we systematically exposed a large number of cells to periodic pulse trains of different frequencies. Our results indicate a resonance peak at a stimulation period of around 20 s. We propose a delayed feedback model that explains our experimental findings based on a time-delay in the regulatory network of the actin system. To test the model, we performed stimulation experiments with cells that express GFP-tagged fusion proteins of Coronin and actin-interacting protein 1, as well as knockout mutants that lack Coronin and actin-interacting protein 1. These actin-binding proteins enhance the disassembly of actin filaments and thus allow us to estimate the delay time in the regulatory feedback loop. Based on this independent estimate, our model predicts an intrinsic period of 20 s, which agrees with the resonance observed in our periodic stimulation experiments. PMID:23431176

  10. Human myosin VIIa is a very slow processive motor protein on various cellular actin structures.

    Science.gov (United States)

    Sato, Osamu; Komatsu, Satoshi; Sakai, Tsuyoshi; Tsukasaki, Yoshikazu; Tanaka, Ryosuke; Mizutani, Takeomi; Watanabe, Tomonobu M; Ikebe, Reiko; Ikebe, Mitsuo

    2017-06-30

    Human myosin VIIa (MYO7A) is an actin-linked motor protein associated with human Usher syndrome (USH) type 1B, which causes human congenital hearing and visual loss. Although it has been thought that the role of human myosin VIIa is critical for USH1 protein tethering with actin and transportation along actin bundles in inner-ear hair cells, myosin VIIa's motor function remains unclear. Here, we studied the motor function of the tail-truncated human myosin VIIa dimer (HM7AΔTail/LZ) at the single-molecule level. We found that the HM7AΔTail/LZ moves processively on single actin filaments with a step size of 35 nm. Dwell-time distribution analysis indicated an average waiting time of 3.4 s, yielding ∼0.3 s -1 for the mechanical turnover rate; hence, the velocity of HM7AΔTail/LZ was extremely slow, at 11 nm·s -1 We also examined HM7AΔTail/LZ movement on various actin structures in demembranated cells. HM7AΔTail/LZ showed unidirectional movement on actin structures at cell edges, such as lamellipodia and filopodia. However, HM7AΔTail/LZ frequently missed steps on actin tracks and exhibited bidirectional movement at stress fibers, which was not observed with tail-truncated myosin Va. These results suggest that the movement of the human myosin VIIa motor protein is more efficient on lamellipodial and filopodial actin tracks than on stress fibers, which are composed of actin filaments with different polarity, and that the actin structures influence the characteristics of cargo transportation by human myosin VIIa. In conclusion, myosin VIIa movement appears to be suitable for translocating USH1 proteins on stereocilia actin bundles in inner-ear hair cells. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  11. Chronologic and actinically induced aging in human facial skin

    International Nuclear Information System (INIS)

    Gilchrest, B.A.; Szabo, G.; Flynn, E.; Goldwyn, R.M.

    1983-01-01

    Clinical and histologic stigmata of aging are much more prominent in habitually sun-exposed skin than in sun-protected skin, but other possible manifestations of actinically induced aging are almost unexplored. We have examined the interrelation of chronologic and actinic aging using paired preauricular (sun-exposed) and postauricular (sun-protected) skin specimens. Keratinocyte cultures derived from sun-exposed skin consistently had a shorter in vitro lifespan but increased plating efficiency compared with cultures derived from adjacent sun-protected skin of the same individual, confirming a previous study of different paired body sites. Electron microscopic histologic sections revealed focal abnormalities of keratinocyte proliferation and alignment in vitro especially in those cultures derived from sun-exposed skin and decreased intercellular contact in stratified colonies at late passage, regardless of donor site. One-micron histologic sections of the original biopsy specimens revealed no striking site-related keratinocyte alterations, but sun-exposed specimens had fewer epidermal Langerhans cells (p less than 0.001), averaging approximately 50 percent the number in sun-protected skin, a possible exaggeration of the previously reported age-associated decrease in this cell population. These data suggest that sun exposure indeed accelerates aging by several criteria and that, regardless of mechanism, environmental factors may adversely affect the appearance and function of aging skin in ways amenable to experimental quantitation

  12. Nano-assembly of nanodiamonds by conjugation to actin filaments.

    Science.gov (United States)

    Bradac, Carlo; Say, Jana M; Rastogi, Ishan D; Cordina, Nicole M; Volz, Thomas; Brown, Louise J

    2016-03-01

    Fluorescent nanodiamonds (NDs) are remarkable objects. They possess unique mechanical and optical properties combined with high surface areas and controllable surface reactivity. They are non-toxic and hence suited for use in biological environments. NDs are also readily available and commercially inexpensive. Here, the exceptional capability of controlling and tailoring their surface chemistry is demonstrated. Small, bright diamond nanocrystals (size ˜30 nm) are conjugated to protein filaments of actin (length ˜3-7 µm). The conjugation to actin filaments is extremely selective and highly target-specific. These unique features, together with the relative simplicity of the conjugation-targeting method, make functionalised nanodiamonds a powerful and versatile platform in biomedicine and quantum nanotechnologies. Applications ranging from using NDs as superior biological markers to, potentially, developing novel bottom-up approaches for the fabrication of hybrid quantum devices that would bridge across the bio/solid-state interface are presented and discussed. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Bacterial actin MreB forms antiparallel double filaments.

    Science.gov (United States)

    van den Ent, Fusinita; Izoré, Thierry; Bharat, Tanmay Am; Johnson, Christopher M; Löwe, Jan

    2014-05-02

    Filaments of all actin-like proteins known to date are assembled from pairs of protofilaments that are arranged in a parallel fashion, generating polarity. In this study, we show that the prokaryotic actin homologue MreB forms pairs of protofilaments that adopt an antiparallel arrangement in vitro and in vivo. We provide an atomic view of antiparallel protofilaments of Caulobacter MreB as apparent from crystal structures. We show that a protofilament doublet is essential for MreB's function in cell shape maintenance and demonstrate by in vivo site-specific cross-linking the antiparallel orientation of MreB protofilaments in E. coli. 3D cryo-EM shows that pairs of protofilaments of Caulobacter MreB tightly bind to membranes. Crystal structures of different nucleotide and polymerisation states of Caulobacter MreB reveal conserved conformational changes accompanying antiparallel filament formation. Finally, the antimicrobial agents A22/MP265 are shown to bind close to the bound nucleotide of MreB, presumably preventing nucleotide hydrolysis and destabilising double protofilaments.DOI: http://dx.doi.org/10.7554/eLife.02634.001. Copyright © 2014, van den Ent et al.

  14. 5-ALA induced fluorescent image analysis of actinic keratosis

    Science.gov (United States)

    Cho, Yong-Jin; Bae, Youngwoo; Choi, Eung-Ho; Jung, Byungjo

    2010-02-01

    In this study, we quantitatively analyzed 5-ALA induced fluorescent images of actinic keratosis using digital fluorescent color and hyperspectral imaging modalities. UV-A was utilized to induce fluorescent images and actinic keratosis (AK) lesions were demarcated from surrounding the normal region with different methods. Eight subjects with AK lesion were participated in this study. In the hyperspectral imaging modality, spectral analysis method was utilized for hyperspectral cube image and AK lesions were demarcated from the normal region. Before image acquisition, we designated biopsy position for histopathology of AK lesion and surrounding normal region. Erythema index (E.I.) values on both regions were calculated from the spectral cube data. Image analysis of subjects resulted in two different groups: the first group with the higher fluorescence signal and E.I. on AK lesion than the normal region; the second group with lower fluorescence signal and without big difference in E.I. between two regions. In fluorescent color image analysis of facial AK, E.I. images were calculated on both normal and AK lesions and compared with the results of hyperspectral imaging modality. The results might indicate that the different intensity of fluorescence and E.I. among the subjects with AK might be interpreted as different phases of morphological and metabolic changes of AK lesions.

  15. Chronic actinic dermatitis - A study of clinical features

    Directory of Open Access Journals (Sweden)

    Somani Vijay

    2005-01-01

    Full Text Available Background: Chronic actinic dermatitis (CAD, one of the immune mediated photo-dermatoses, comprises a spectrum of conditions including persistent light reactivity, photosensitive eczema and actinic reticuloid. Diagnostic criteria were laid down about 20 years back, but clinical features are the mainstay in diagnosis. In addition to extreme sensitivity to UVB, UVA and/or visible light, about three quarters of patients exhibit contact sensitivity to several allergens, which may contribute to the etiopathogenesis of CAD. This study was undertaken to examine the clinical features of CAD in India and to evaluate the relevance of patch testing and photo-aggravation testing in the diagnosis of CAD. Methods: The clinical data of nine patients with CAD were analyzed. Histopathology, patch testing and photo-aggravation testing were also performed. Results: All the patients were males. The average age of onset was 57 years. The first episode was usually noticed in the beginning of summer. Later the disease gradually tended to be perennial, without any seasonal variations. The areas affected were mainly the photo-exposed areas in all patients, and the back in three patients. Erythroderma was the presenting feature in two patients. The palms and soles were involved in five patients. Patch testing was positive in seven of nine patients. Conclusions: The diagnosis of CAD mainly depended upon the history and clinical features. The incidence of erythroderma and palmoplantar eczema was high in our series. Occupation seems to play a role in the etiopathogenesis of CAD.

  16. Valoración de la superficie del dializador en la hemodiafiltración on-line. Elección objetiva de la superficie del dializador

    Directory of Open Access Journals (Sweden)

    Francisco Maduell

    2015-05-01

    Conclusión: El incremento del 40% y el 80% de la superficie conlleva un aumento del volumen convectivo de un 6 y un 16% respectivamente, aunque se evidenció una reducción en su máximo rendimiento, mostrando mínimas diferencias tanto en el volumen convectivo como en la capacidad depurativa cuando el CUF era superior a 45 ml/h/mmHg. Es recomendable optimizar el rendimiento de los dializadores a la mínima superficie posible adecuando la prescripción de tratamiento.

  17. The Stationary-Phase Cells of Saccharomyces cerevisiae Display Dynamic Actin Filaments Required for Processes Extending Chronological Life Span.

    Science.gov (United States)

    Vasicova, Pavla; Lejskova, Renata; Malcova, Ivana; Hasek, Jiri

    2015-11-01

    Stationary-growth-phase Saccharomyces cerevisiae yeast cultures consist of nondividing cells that undergo chronological aging. For their successful survival, the turnover of proteins and organelles, ensured by autophagy and the activation of mitochondria, is performed. Some of these processes are engaged in by the actin cytoskeleton. In S. cerevisiae stationary-phase cells, F actin has been shown to form static aggregates named actin bodies, subsequently cited to be markers of quiescence. Our in vivo analyses revealed that stationary-phase cultures contain cells with dynamic actin filaments, besides the cells with static actin bodies. The cells with dynamic actin displayed active endocytosis and autophagy and well-developed mitochondrial networks. Even more, stationary-phase cell cultures grown under calorie restriction predominantly contained cells with actin cables, confirming that the presence of actin cables is linked to successful adaptation to stationary phase. Cells with actin bodies were inactive in endocytosis and autophagy and displayed aberrations in mitochondrial networks. Notably, cells of the respiratory activity-deficient cox4Δ strain displayed the same mitochondrial aberrations and actin bodies only. Additionally, our results indicate that mitochondrial dysfunction precedes the formation of actin bodies and the appearance of actin bodies corresponds to decreased cell fitness. We conclude that the F-actin status reflects the extent of damage that arises from exponential growth. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  18. Roles of Asp179 and Glu270 in ADP-Ribosylation of Actin by Clostridium perfringens Iota Toxin.

    Directory of Open Access Journals (Sweden)

    Alexander Belyy

    Full Text Available Clostridium perfringens iota toxin is a binary toxin composed of the enzymatically active component Ia and receptor binding component Ib. Ia is an ADP-ribosyltransferase, which modifies Arg177 of actin. The previously determined crystal structure of the actin-Ia complex suggested involvement of Asp179 of actin in the ADP-ribosylation reaction. To gain more insights into the structural requirements of actin to serve as a substrate for toxin-catalyzed ADP-ribosylation, we engineered Saccharomyces cerevisiae strains, in which wild type actin was replaced by actin variants with substitutions in residues located on the Ia-actin interface. Expression of the actin mutant Arg177Lys resulted in complete resistance towards Ia. Actin mutation of Asp179 did not change Ia-induced ADP-ribosylation and growth inhibition of S. cerevisiae. By contrast, substitution of Glu270 of actin inhibited the toxic action of Ia and the ADP-ribosylation of actin. In vitro transcribed/translated human β-actin confirmed the crucial role of Glu270 in ADP-ribosylation of actin by Ia.

  19. Axon initial segment cytoskeleton comprises a multiprotein submembranous coat containing sparse actin filaments

    Science.gov (United States)

    Jones, Steven L.; Korobova, Farida

    2014-01-01

    The axon initial segment (AIS) of differentiated neurons regulates action potential initiation and axon–dendritic polarity. The latter function depends on actin dynamics, but actin structure and functions at the AIS remain unclear. Using platinum replica electron microscopy (PREM), we have characterized the architecture of the AIS cytoskeleton in mature and developing hippocampal neurons. The AIS cytoskeleton assembly begins with bundling of microtubules and culminates in formation of a dense, fibrillar–globular coat over microtubule bundles. Immunogold PREM revealed that the coat contains a network of known AIS proteins, including ankyrin G, spectrin βIV, neurofascin, neuronal cell adhesion molecule, voltage-gated sodium channels, and actin filaments. Contrary to existing models, we find neither polarized actin arrays, nor dense actin meshworks in the AIS. Instead, the AIS contains two populations of sparse actin filaments: short, stable filaments and slightly longer dynamic filaments. We propose that stable actin filaments play a structural role for formation of the AIS diffusion barrier, whereas dynamic actin may promote AIS coat remodeling. PMID:24711503

  20. Actin and myosin regulate cytoplasm stiffness in plant cells: a study using optical tweezers

    NARCIS (Netherlands)

    Honing, van der H.S.; Ruijter, de N.C.A.; Emons, A.M.C.; Ketelaar, T.

    2010-01-01

    Here, we produced cytoplasmic protrusions with optical tweezers in mature BY-2 suspension cultured cells to study the parameters involved in the movement of actin filaments during changes in cytoplasmic organization and to determine whether stiffness is an actin-related property of plant cytoplasm.

  1. Actin based processes that could determine the cytoplasmic architecture of plant cells

    NARCIS (Netherlands)

    Honing, van der H.S.; Emons, A.M.C.; Ketelaar, M.J.

    2007-01-01

    Actin polymerisation can generate forces that are necessary for cell movement, such as the propulsion of a class of bacteria, including Listeria, and the protrusion of migrating animal cells. Force generation by the actin cytoskeleton in plant cells has not been studied. One process in plant cells

  2. Allyl Isothiocyanate Inhibits Actin-Dependent Intracellular Transport in Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Bjørnar Sporsheim

    2015-12-01

    Full Text Available Volatile allyl isothiocyanate (AITC derives from the biodegradation of the glucosinolate sinigrin and has been associated with growth inhibition in several plants, including the model plant Arabidopsis thaliana. However, the underlying cellular mechanisms of this feature remain scarcely investigated in plants. In this study, we present evidence of an AITC-induced inhibition of actin-dependent intracellular transport in A. thaliana. A transgenic line of A. thaliana expressing yellow fluorescent protein (YFP-tagged actin filaments was used to show attenuation of actin filament movement by AITC. This appeared gradually in a time- and dose-dependent manner and resulted in actin filaments appearing close to static. Further, we employed four transgenic lines with YFP-fusion proteins labeling the Golgi apparatus, endoplasmic reticulum (ER, vacuoles and peroxisomes to demonstrate an AITC-induced inhibition of actin-dependent intracellular transport of or, in these structures, consistent with the decline in actin filament movement. Furthermore, the morphologies of actin filaments, ER and vacuoles appeared aberrant following AITC-exposure. However, AITC-treated seedlings of all transgenic lines tested displayed morphologies and intracellular movements similar to that of the corresponding untreated and control-treated plants, following overnight incubation in an AITC-absent environment, indicating that AITC-induced decline in actin-related movements is a reversible process. These findings provide novel insights into the cellular events in plant cells following exposure to AITC, which may further expose clues to the physiological significance of the glucosinolate-myrosinase system.

  3. When fat is not bad: the regulation of actin dynamics by phospholipid signaling molecules

    Czech Academy of Sciences Publication Activity Database

    Pleskot, Roman; Pejchar, Přemysl; Staiger, Ch. J.; Potocký, Martin

    2014-01-01

    Roč. 5, JAN 2014 (2014) ISSN 1664-462X R&D Projects: GA ČR GA13-19073S Institutional support: RVO:61389030 Keywords : actin * actin-binding proteins * capping protein Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.948, year: 2014

  4. The actin homologue MreB organizes the bacterial cell membrane

    NARCIS (Netherlands)

    Strahl, H.; Burmann, F.; Hamoen, L.W.

    2014-01-01

    The eukaryotic cortical actin cytoskeleton creates specific lipid domains, including lipid rafts, which determine the distribution of many membrane proteins. Here we show that the bacterial actin homologue MreB displays a comparable activity. MreB forms membrane-associated filaments that coordinate

  5. The unique organization of filamentous actin in the medullary canal of the medulla oblongata.

    Science.gov (United States)

    Tan, Bai-Hong; Guo, Chun-Yan; Xiong, Tian-Qing; Chen, Ling-Meng; Li, Yan-Chao

    2017-04-01

    In the central canal, F-actin is predominantly localized in the apical region, forming a ring-like structure around the circumference of the lumen. However, an exception is found in the medulla oblongata, where the apical F-actin becomes interrupted in the ventral aspect of the canal. To clarify the precise localization of F-actin, the fluorescence signals for F-actin were converted to the peroxidase/DAB reaction products in this study by a phalloidin-based ultrastructural technique, which demonstrated that F-actin is located mainly in the microvilli and terminal webs in the ependymocytes. It is because the ventrally oriented ependymocytes do not possess well-developed microvilli or terminal web that led to a discontinuous labeling of F-actin in the medullary canal. Since spinal motions can change the shape and size of the central canal, we next examined the cytoskeletons in the medullary canal in both rats and monkeys, because these two kinds of animals show different kinematics at the atlanto-occipital articulation. Our results first demonstrated that the apical F-actin in the medullary canal is differently organized in the animals with different head-neck kinemics, which suggests that the mechanic stretching of spinal motions is capable of inducing F-actin reorganization and the subsequent cell-shape changes in the central canal. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. Changes in actin dynamics are involved in salicylic acid signaling pathway.

    Science.gov (United States)

    Matoušková, Jindřiška; Janda, Martin; Fišer, Radovan; Sašek, Vladimír; Kocourková, Daniela; Burketová, Lenka; Dušková, Jiřina; Martinec, Jan; Valentová, Olga

    2014-06-01

    Changes in actin cytoskeleton dynamics are one of the crucial players in many physiological as well as non-physiological processes in plant cells. Positioning of actin filament arrays is necessary for successful establishment of primary lines of defense toward pathogen attack, depolymerization leads very often to the enhanced susceptibility to the invading pathogen. On the other hand it was also shown that the disruption of actin cytoskeleton leads to the induction of defense response leading to the expression of PATHOGENESIS RELATED proteins (PR). In this study we show that pharmacological actin depolymerization leads to the specific induction of genes in salicylic acid pathway but not that involved in jasmonic acid signaling. Life imaging of leafs of Arabidopsis thaliana with GFP-tagged fimbrin (GFP-fABD2) treated with 1 mM salicylic acid revealed rapid disruption of actin filaments resembling the pattern viewed after treatment with 200 nM latrunculin B. The effect of salicylic acid on actin filament fragmentation was prevented by exogenous addition of phosphatidic acid, which binds to the capping protein and thus promotes actin polymerization. The quantitative evaluation of actin filament dynamics is also presented. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  7. Disruption of microtubule network rescues aberrant actin comets in dynamin2-depleted cells.

    Directory of Open Access Journals (Sweden)

    Yuji Henmi

    Full Text Available A large GTPase dynamin, which is required for endocytic vesicle formation, regulates the actin cytoskeleton through its interaction with cortactin. Dynamin2 mutants impair the formation of actin comets, which are induced by Listeria monocytogenes or phosphatidylinositol-4-phosphate 5-kinase. However, the role of dynamin2 in the regulation of the actin comet is still unclear. Here we show that aberrant actin comets in dynamin2-depleted cells were rescued by disrupting of microtubule networks. Depletion of dynamin2, but not cortactin, significantly reduced the length and the speed of actin comets induced by Listeria. This implies that dynamin2 may regulate the actin comet in a cortactin-independent manner. As dynamin regulates microtubules, we investigated whether perturbation of microtubules would rescue actin comet formation in dynamin2-depleted cells. Treatment with taxol or colchicine created a microtubule-free space in the cytoplasm, and made no difference between control and dynamin2 siRNA cells. This suggests that the alteration of microtubules by dynamin2 depletion reduced the length and the speed of the actin comet.

  8. Total Synthesis of (-)-Doliculide, Structure-Activity Relationship Studies and Its Binding to F-Actin

    NARCIS (Netherlands)

    Matcha, Kiran; Madduri, Ashoka V. R.; Roy, Sayantani; Ziegler, Slava; Waldmann, Herbert; Hirsch, Anna K. H.; Minnaard, Adriaan J.

    2012-01-01

    Actin, an abundant protein in most eukaryotic cells, is one of the targets in cancer research. Recently, a great deal of attention has been paid to the synthesis and function of actin-targeting compounds and their use as effective molecular probes in chemical biology. In this study, we have

  9. The Actin-Binding Protein α-Adducin Is Required for Maintaining Axon Diameter

    Directory of Open Access Journals (Sweden)

    Sérgio Carvalho Leite

    2016-04-01

    Full Text Available The actin-binding protein adducin was recently identified as a component of the neuronal subcortical cytoskeleton. Here, we analyzed mice lacking adducin to uncover the function of this protein in actin rings. α-adducin knockout mice presented progressive axon enlargement in the spinal cord and optic and sciatic nerves, followed by axon degeneration and loss. Using stimulated emission depletion super-resolution microscopy, we show that a periodic subcortical actin cytoskeleton is assembled in every neuron type inspected including retinal ganglion cells and dorsal root ganglia neurons. In neurons devoid of adducin, the actin ring diameter increased, although the inter-ring periodicity was maintained. In vitro, the actin ring diameter adjusted as axons grew, suggesting the lattice is dynamic. Our data support a model in which adducin activity is not essential for actin ring assembly and periodicity but is necessary to control the diameter of both actin rings and axons and actin filament growth within rings.

  10. Axonal regeneration and neuronal function are preserved in motor neurons lacking ß-actin in vivo.

    Directory of Open Access Journals (Sweden)

    Thomas R Cheever

    2011-03-01

    Full Text Available The proper localization of ß-actin mRNA and protein is essential for growth cone guidance and axon elongation in cultured neurons. In addition, decreased levels of ß-actin mRNA and protein have been identified in the growth cones of motor neurons cultured from a mouse model of Spinal Muscular Atrophy (SMA, suggesting that ß-actin loss-of-function at growth cones or pre-synaptic nerve terminals could contribute to the pathogenesis of this disease. However, the role of ß-actin in motor neurons in vivo and its potential relevance to disease has yet to be examined. We therefore generated motor neuron specific ß-actin knock-out mice (Actb-MNsKO to investigate the function of ß-actin in motor neurons in vivo. Surprisingly, ß-actin was not required for motor neuron viability or neuromuscular junction maintenance. Skeletal muscle from Actb-MNsKO mice showed no histological indication of denervation and did not significantly differ from controls in several measurements of physiologic function. Finally, motor axon regeneration was unimpaired in Actb-MNsKO mice, suggesting that ß-actin is not required for motor neuron function or regeneration in vivo.

  11. Disassembly of actin structures by nanosecond pulsed electric field is a downstream effect of cell swelling.

    Science.gov (United States)

    Pakhomov, Andrei G; Xiao, Shu; Pakhomova, Olga N; Semenov, Iurii; Kuipers, Marjorie A; Ibey, Bennett L

    2014-12-01

    Disruption of the actin cytoskeleton structures was reported as one of the characteristic effects of nanosecond-duration pulsed electric field (nsPEF) in both mammalian and plant cells. We utilized CHO cells that expressed the monomeric fluorescent protein (mApple) tagged to actin to test if nsPEF modifies the cell actin directly or as a consequence of cell membrane permeabilization. A train of four 600-ns pulses at 19.2 kV/cm (2 Hz) caused immediate cell membrane poration manifested by YO-PRO-1 dye uptake, gradual cell rounding and swelling. Concurrently, bright actin features were replaced by dimmer and uniform fluorescence of diffuse actin. To block the nsPEF-induced swelling, the bath buffer was isoosmotically supplemented with an electropore-impermeable solute (sucrose). A similar addition of a smaller, electropore-permeable solute (adonitol) served as a control. We demonstrated that sucrose efficiently blocked disassembly of actin features by nsPEF, whereas adonitol did not. Sucrose also attenuated bleaching of mApple-tagged actin in nsPEF-treated cells (as integrated over the cell volume), although did not fully prevent it. We conclude that disintegration of the actin cytoskeleton was a result of cell swelling, which, in turn, was caused by cell permeabilization by nsPEF and transmembrane diffusion of solutes which led to the osmotic imbalance. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Regulation of the actin cytoskeleton-plasma membrane interplay by phosphoinositides.

    Science.gov (United States)

    Saarikangas, Juha; Zhao, Hongxia; Lappalainen, Pekka

    2010-01-01

    The plasma membrane and the underlying cortical actin cytoskeleton undergo continuous dynamic interplay that is responsible for many essential aspects of cell physiology. Polymerization of actin filaments against cellular membranes provides the force for a number of cellular processes such as migration, morphogenesis, and endocytosis. Plasma membrane phosphoinositides (especially phosphatidylinositol bis- and trisphosphates) play a central role in regulating the organization and dynamics of the actin cytoskeleton by acting as platforms for protein recruitment, by triggering signaling cascades, and by directly regulating the activities of actin-binding proteins. Furthermore, a number of actin-associated proteins, such as BAR domain proteins, are capable of directly deforming phosphoinositide-rich membranes to induce plasma membrane protrusions or invaginations. Recent studies have also provided evidence that the actin cytoskeleton-plasma membrane interactions are misregulated in a number of pathological conditions such as cancer and during pathogen invasion. Here, we summarize the wealth of knowledge on how the cortical actin cytoskeleton is regulated by phosphoinositides during various cell biological processes. We also discuss the mechanisms by which interplay between actin dynamics and certain membrane deforming proteins regulate the morphology of the plasma membrane.

  13. NHERF1 regulates actin cytoskeleton organization through modulation of α-actinin-4 stability.

    Science.gov (United States)

    Sun, Licui; Zheng, Junfang; Wang, Qiqi; Song, Ran; Liu, Hua; Meng, Ran; Tao, Tao; Si, Yang; Jiang, Wenguo; He, Junqi

    2016-02-01

    The actin cytoskeleton is composed of a highly dynamic network of filamentous proteins, yet the molecular mechanism that regulates its organization and remodeling remains elusive. In this study, Na(+)/H(+) exchanger regulatory factor (NHERF)-1 loss-of-function and gain-of-function experiments reveal that polymerized actin cytoskeleton (F-actin) in HeLa cells is disorganized by NHERF1, whereas actin protein expression levels exhibit no detectable change. To elucidate the molecular mechanism underlying actin cytoskeleton disorganization by NHERF1, a combined 2-dimensional electrophoresis-matrix-assisted laser desorption/ionization-time of flight mass spectrometry approach was used to screen for proteins regulated by NHERF1 in HeLa cells. α-Actinin-4, an actin cross-linking protein, was identified. Glutathione S-transferase pull-down and coimmunoprecipitation studies showed the α-actinin-4 carboxyl-terminal region specifically interacted with the NHERF1 postsynaptic density 95/disc-large/zona occludens-1 domain. The NHERF1/α-actinin-4 interaction increased α-actinin-4 ubiquitination and decreased its expression levels, resulting in actin cytoskeleton disassembly. Our study identified α-actinin-4 as a novel NHERF1 interaction partner and provided new insights into the regulatory mechanism of the actin cytoskeleton by NHERF1. © FASEB.

  14. Impaired recycling of synaptic vesicles after acute perturbation of the presynaptic actin cytoskeleton

    DEFF Research Database (Denmark)

    Shupliakov, Oleg; Bloom, Ona; Gustafsson, Jenny S

    2002-01-01

    Actin is an abundant component of nerve terminals that has been implicated at multiple steps of the synaptic vesicle cycle, including reversible anchoring, exocytosis, and recycling of synaptic vesicles. In the present study we used the lamprey reticulospinal synapse to examine the role of actin ...

  15. The Actin-Binding Protein α-Adducin Is Required for Maintaining Axon Diameter.

    Science.gov (United States)

    Leite, Sérgio Carvalho; Sampaio, Paula; Sousa, Vera Filipe; Nogueira-Rodrigues, Joana; Pinto-Costa, Rita; Peters, Luanne Laurel; Brites, Pedro; Sousa, Mónica Mendes

    2016-04-19

    The actin-binding protein adducin was recently identified as a component of the neuronal subcortical cytoskeleton. Here, we analyzed mice lacking adducin to uncover the function of this protein in actin rings. α-adducin knockout mice presented progressive axon enlargement in the spinal cord and optic and sciatic nerves, followed by axon degeneration and loss. Using stimulated emission depletion super-resolution microscopy, we show that a periodic subcortical actin cytoskeleton is assembled in every neuron type inspected including retinal ganglion cells and dorsal root ganglia neurons. In neurons devoid of adducin, the actin ring diameter increased, although the inter-ring periodicity was maintained. In vitro, the actin ring diameter adjusted as axons grew, suggesting the lattice is dynamic. Our data support a model in which adducin activity is not essential for actin ring assembly and periodicity but is necessary to control the diameter of both actin rings and axons and actin filament growth within rings. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  16. Rapid and dynamic arginylation of the leading edge β-actin is required for cell migration.

    Science.gov (United States)

    Pavlyk, Iuliia; Leu, Nicolae A; Vedula, Pavan; Kurosaka, Satoshi; Kashina, Anna

    2018-04-01

    β-actin plays key roles in cell migration. Our previous work demonstrated that β-actin in migratory non-muscle cells is N-terminally arginylated and that this arginylation is required for normal lamellipodia extension. Here, we examined the function of β-actin arginylation in cell migration. We found that arginylated β-actin is concentrated at the leading edge of lamellipodia and that this enrichment is abolished after serum starvation as well as in contact-inhibited cells in confluent cultures, suggesting that arginylated β-actin at the cell leading edge is coupled to active migration. Arginylated actin levels exhibit dynamic changes in response to cell stimuli, lowered after serum starvation and dramatically elevating within minutes after cell stimulation by readdition of serum or lysophosphatidic acid. These dynamic changes require active translation and are not seen in confluent contact-inhibited cell cultures. Microinjection of arginylated actin antibodies into cells severely and specifically inhibits their migration rates. Together, these data strongly suggest that arginylation of β-actin is a tightly regulated dynamic process that occurs at the leading edge of locomoting cells in response to stimuli and is integral to the signaling network that regulates cell migration. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  17. Plasticity of the actin cytoskeleton in response to extracellular matrix nanostructure and dimensionality

    NARCIS (Netherlands)

    Starke, J.; Wehrle-Haller, B.; Friedl, P.

    2014-01-01

    Mobile cells discriminate and adapt to mechanosensory input from extracellular matrix (ECM) topographies to undergo actin-based polarization, shape change and migration. We tested 'cell-intrinsic' and adaptive components of actin-based cell migration in response to widely used in vitro

  18. Biomass torrefaction characteristics in inert and oxidative atmospheres at various superficial velocities.

    Science.gov (United States)

    Chen, Wei-Hsin; Lu, Ke-Miao; Liu, Shih-Hsien; Tsai, Chi-Ming; Lee, Wen-Jhy; Lin, Ta-Chang

    2013-10-01

    The reaction characteristics of four biomass materials (i.e. oil palm fiber, coconut fiber, eucalyptus, and Cryptomeria japonica) with non-oxidative and oxidative torrefaction at various superficial velocities are investigated where nitrogen and air are used as carrier gases. Three torrefaction temperatures of 250, 300, and 350 °C are considered. At a given temperature, the solid yield of biomass is not affected by N2 superficial velocity, revealing that the thermal degradation is controlled by heat and mass transfer in biomass. Increasing air superficial velocity decreases the solid yield, especially in oil palm fiber and coconut fiber, implying that the torrefaction reaction of biomass is dominated by surface oxidation. There exists an upper limit of air superficial velocity in the decrement of solid yield, suggesting that beyond this limit the thermal degradation of biomass is no longer governed by surface oxidation, but rather is controlled by internal mass transport. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. Physiological Laterality of Superficial Cerebral Veins on Susceptibility-Weighted Imaging.

    Science.gov (United States)

    Matsushima, Satoshi; Shimizu, Tetsuya; Gomi, Taku; Fukuda, Kunihiko

    The purpose of this study is to evaluate whether laterality of the superficial cerebral veins can be seen on susceptibility-weighted imaging (SWI) in patients with no intracranial lesions that affect venous visualization. We retrospectively evaluated 386 patients who underwent brain magnetic resonance imaging including SWI in our institute. Patients with a lesion with the potential to affect venous visualization on SWI were excluded. Two neuroradiologists visually evaluated the findings and scored the visualization of the superficial cerebral veins. Of the 386 patients, 315 (81.6%) showed no obvious laterality on venous visualization, 64 (16.6%) showed left-side dominant laterality, and 7 (1.8%) showed right-side dominant laterality. Left-side dominant physiological laterality exists in the visualization of the superficial cerebral veins on SWI. Therefore, when recognizing left-side dominant laterality of the superficial cerebral veins on SWI, the radiologist must also consider the possibility of physiological laterality.

  20. DETERMINATION OF SUPERFICIAL ABSORBED DOSE FROM EXTERNAL EXPOSURE OF WEAKLY PENETRATING RADIATIONS

    Institute of Scientific and Technical Information of China (English)

    陈丽姝

    1994-01-01

    The methods of determining the superficial absorbed dose distributions in a water phantom by means of the experiments and available theories have been reported.The distributions of beta dose were measured by an extrapolation ionization chamber at definite depthes corresponding to some superficial organs and tissues such as the radiosensitive layer of the skin,cornea,sclera,anterior chamber and lens of eyeball.The ratios among superficial absorbed dose D(0.07) and average absorbed doses at the depthes 1,2,3,4,5 and 6mm are also obtained with Cross's methods.They can be used for confining the deterministic effects of some superficial tissues and organs such as the skin and the components of eyeball for weakly penetrating radiations.

  1. Effects of immobilization on thickness of superficial zone of articular cartilage of patella in rats

    Directory of Open Access Journals (Sweden)

    Khadija Iqbal

    2012-01-01

    Conclusion: Each segment of superficial zone behaves differentially on immobilization and remobilization. Perhaps a much longer duration of remobilization is required to reverse changes of immobilization in articular cartilage and plays a significant role in knee joint movements.

  2. Terbinafine-loaded wound dressing for chronic superficial fungal infections

    International Nuclear Information System (INIS)

    Paskiabi, Farnoush Asghari; Bonakdar, Shahin; Shokrgozar, Mohammad Ali; Imani, Mohammad; Jahanshiri, Zahra; Shams-Ghahfarokhi, Masoomeh; Razzaghi-Abyaneh, Mehdi

    2017-01-01

    In spite of developing new drugs and modern formulations, the treatments of chronic fungal infections are still challenging. Fibrous wound dressings are new suggestions for the treatment of chronic superficial infections. In the present study, we formulated an antifungal agent, terbinafine hydrochloride (TFH), which is a hydrophobic drug, in wound dressings prepared by electrospun polycaprolactone, polycaprolactone/gelatin (50:50 w/w) and gelatin. To obtain more water-stable meshes, the preparations were treated by glutaraldehyde and their properties were determined before and after treatment. The morphology of fibrous meshes was observed by scanning electron microscopy. Drug loading efficiency and release rate were measured by high performance liquid chromatography (HPLC) and the release rate was monitored for 144 h. Antifungal tests were performed on Trichophyton mentagrophytes, Aspergillus fumigatus and Candida albicans cultured on Muller-Hinton agar. The toxicity of the meshes was measured after 24 h and 14 days by MTT assay. Terbinafine loading of polycaprolactone/gelatin (50:50) was 100% and it released the highest amount of TFH too. In antifungal tests, all samples were able to hinderT. mentagrophytes and A. fumigatus but not C. albicans growth among them, polycaprolactone fibers made the largest inhibition zone. In MTT assay, none of prepared samples showed toxicity against L929 cells. Teken together, the prepared TFH-loaded PCL/gelatin electrospun meshes were able to release TFH slowly and in a steady state in time. With respect to no obvious cytotoxicity in MTT assay and stong antifungal activity toward T. mentagrophytesin vitro, these TFH-based meshes could be considered as potential candidates in clinical application as wound dressing for treatment of chronic dermatophytosis. - Highlights: • Terbinafine (TFH)-loaded PCL/gelatin electrospun fibers were successfully fabricated. • TFH-loaded PCL/gelatin electrospun fibers showed a slow drug release

  3. Terbinafine-loaded wound dressing for chronic superficial fungal infections

    Energy Technology Data Exchange (ETDEWEB)

    Paskiabi, Farnoush Asghari [Department of Mycology, Pasteur Institute of Iran, Tehran 13164. Iran (Iran, Islamic Republic of); Microbiology Research Center, Pasteur Institute of Iran, Tehran 13164. Iran (Iran, Islamic Republic of); Bonakdar, Shahin; Shokrgozar, Mohammad Ali [National Cell Bank Department, Pasteur Institute of Iran, Tehran 13164 (Iran, Islamic Republic of); Imani, Mohammad [Department of Novel Drug Delivery Systems, Iran Polymer and Petrochemical Institute, Tehran (Iran, Islamic Republic of); Jahanshiri, Zahra [Department of Mycology, Pasteur Institute of Iran, Tehran 13164. Iran (Iran, Islamic Republic of); Shams-Ghahfarokhi, Masoomeh [Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran (Iran, Islamic Republic of); Razzaghi-Abyaneh, Mehdi, E-mail: mrab442@yahoo.com [Department of Mycology, Pasteur Institute of Iran, Tehran 13164. Iran (Iran, Islamic Republic of); Microbiology Research Center, Pasteur Institute of Iran, Tehran 13164. Iran (Iran, Islamic Republic of)

    2017-04-01

    In spite of developing new drugs and modern formulations, the treatments of chronic fungal infections are still challenging. Fibrous wound dressings are new suggestions for the treatment of chronic superficial infections. In the present study, we formulated an antifungal agent, terbinafine hydrochloride (TFH), which is a hydrophobic drug, in wound dressings prepared by electrospun polycaprolactone, polycaprolactone/gelatin (50:50 w/w) and gelatin. To obtain more water-stable meshes, the preparations were treated by glutaraldehyde and their properties were determined before and after treatment. The morphology of fibrous meshes was observed by scanning electron microscopy. Drug loading efficiency and release rate were measured by high performance liquid chromatography (HPLC) and the release rate was monitored for 144 h. Antifungal tests were performed on Trichophyton mentagrophytes, Aspergillus fumigatus and Candida albicans cultured on Muller-Hinton agar. The toxicity of the meshes was measured after 24 h and 14 days by MTT assay. Terbinafine loading of polycaprolactone/gelatin (50:50) was 100% and it released the highest amount of TFH too. In antifungal tests, all samples were able to hinderT. mentagrophytes and A. fumigatus but not C. albicans growth among them, polycaprolactone fibers made the largest inhibition zone. In MTT assay, none of prepared samples showed toxicity against L929 cells. Teken together, the prepared TFH-loaded PCL/gelatin electrospun meshes were able to release TFH slowly and in a steady state in time. With respect to no obvious cytotoxicity in MTT assay and stong antifungal activity toward T. mentagrophytesin vitro, these TFH-based meshes could be considered as potential candidates in clinical application as wound dressing for treatment of chronic dermatophytosis. - Highlights: • Terbinafine (TFH)-loaded PCL/gelatin electrospun fibers were successfully fabricated. • TFH-loaded PCL/gelatin electrospun fibers showed a slow drug release

  4. Radiotherapy for superficial esophageal cancer of poor risk patients

    International Nuclear Information System (INIS)

    Kagami, Yoshikazu; Ikeda, Hiroshi; Murayama, Shigeyuki; Yamaguchi, Hajime; Tachimori, Yuji; Kato, Hoichi; Watanabe, Hiroshi; Tokuue, Kouichi; Sumi, Minako; Kawashima, Mitsuhiko; Imai, Atsushi; Nakayama, Shuji

    1997-01-01

    Purpose/Objective: The reported incidence of superficial esophageal cancer (SEC) has steadily increased in Japan as result of endoscopic examination has been become common. In Japan, treatment of SEC is endoscopical mucosal resection (EMR) for mucosal cancer or esophagectomy with 3 fields lymph nodes resection for submucosal cancer. Radiotherapy is little place for the management of SEC. Because of some reasons, we treated patients with SEC by radiotherapy alternative to surgery. Purpose of this report is to evaluate efficacy of radiotherapy for SEC. Methods and Materials: Between 1989 to 1996, eighteen patients with SEC were treated with radiotherapy at our hospital. Reasons of radiotherapy that was chosen as the primary methods of treatment were refusal of surgery in one patient, poor medical condition in 4 patients and double primary cancer in 13 patients (head and neck: 11, simultaneously: 11). No patients had indication of EMR. Diagnosis was made by endoscopy and radiography. Some patients were examined with endoscopic ultrasound. Two patients (11.1%) had tumor limited to the mucosa and 16 patients (88.9%) had tumor invaded the submucosa. Seven of these tumors (38.9%) were multicentric. All patients had squamous cell carcinoma. There were 17 male patients and one female patient. The age range was 49 years to 87 years with a median of 62 years. Stage of all patients was T1N0M0 according to UICC staging system. Ten patients underwent external radiotherapy (Ex) (50 Gy - 66 Gy) alone and 8 patients did both Ex and intracavitary radiotherapy (IC) (30-60 Gy of Ex with 5-15 Gy of IC). No patients received chemotherapy. Duration of follow-up was 6 months to 96 months with a median of 30 months. Results: The overall survival rate was 55.9% in 3-year and 14% in 5-year, and the cause-specific 5-year survival rate was 100%. Causes of death were malignant tumor other than esophageal cancer in 4 patients, intercurrent disease other than malignant tumor in 3 patients and no

  5. Interaction of mobile phones with superficial passive metallic implants

    International Nuclear Information System (INIS)

    Virtanen, H; Huttunen, J; Toropainen, A; Lappalainen, R

    2005-01-01

    The dosimetry of exposure to radiofrequency (RF) electromagnetic (EM) fields of mobile phones is generally based on the specific absorption rate (SAR, W kg -1 ), which is the electromagnetic energy absorbed in the tissues per unit mass and time. In this study, numerical methods and modelling were used to estimate the effect of a passive, metallic (conducting) superficial implant on a mobile phone EM field and especially its absorption in tissues in the near field. Two basic implant models were studied: metallic pins and rings in the surface layers of the human body near the mobile phone. The aim was to find out 'the worst case scenario' with respect to energy absorption by varying different parameters such as implant location, orientation, size and adjacent tissues. Modelling and electromagnetic field calculations were carried out using commercial SEMCAD software based on the FDTD (finite difference time domain) method. The mobile phone was a 900 MHz or 1800 MHz generic phone with a quarter wave monopole antenna. A cylindrical tissue phantom models different curved sections of the human body such as limbs or a head. All the parameters studied (implant size, orientation, location, adjacent tissues and signal frequency) had a major effect on the SAR distribution and in certain cases high local EM fields arose near the implant. The SAR values increased most when the implant was on the skin and had a resonance length or diameter, i.e. about a third of the wavelength in tissues. The local peak SAR values increased even by a factor of 400-700 due to a pin or a ring. These highest values were reached in a limited volume close to the implant surface in almost all the studied cases. In contrast, without the implant the highest SAR values were generally reached on the skin surface. Mass averaged SAR 1g and SAR 10g values increased due to the implant even by a factor of 3 and 2, respectively. However, at typical power levels of mobile phones the enhancement is unlikely to be

  6. A single charge in the actin binding domain of fascin can independently tune the linear and non-linear response of an actin bundle network.

    Science.gov (United States)

    Maier, M; Müller, K W; Heussinger, C; Köhler, S; Wall, W A; Bausch, A R; Lieleg, O

    2015-05-01

    Actin binding proteins (ABPs) not only set the structure of actin filament assemblies but also mediate the frequency-dependent viscoelastic moduli of cross-linked and bundled actin networks. Point mutations in the actin binding domain of those ABPs can tune the association and dissociation dynamics of the actin/ABP bond and thus modulate the network mechanics both in the linear and non-linear response regime. We here demonstrate how the exchange of a single charged amino acid in the actin binding domain of the ABP fascin triggers such a modulation of the network rheology. Whereas the overall structure of the bundle networks is conserved, the transition point from strain-hardening to strain-weakening sensitively depends on the cross-linker off-rate and the applied shear rate. Our experimental results are consistent both with numerical simulations of a cross-linked bundle network and a theoretical description of the bundle network mechanics which is based on non-affine bending deformations and force-dependent cross-link dynamics.

  7. A Novel Technique of Supra Superficial Musculoaponeurotic System Hyaluronic Acid Injection for Lower Face Lifting

    OpenAIRE

    Sahawatwong, Sinijchaya; Sirithanabadeekul, Punyaphat; Patanajareet, Vasiyapha; Wattanakrai, Penpun; Thanasarnaksorn, Wilai

    2016-01-01

    Background: Various methods attempting to correct sagging of the lower face focus mainly on manipulation of the superficial musculoaponeurotic System. Each technique has its own limitation. The authors propose a relatively simple, conservative method utilizing hyaluronic acid injection just above the superficial musculoaponeurotic System. Objective: To address a novel hyaluronic injection technique to lift the lower face. Methods: Details of the injection techniques are described. The Positio...

  8. Clinicomycological Characterization of Superficial Mycoses from a Tertiary Care Hospital in Nepal

    OpenAIRE

    Khadka, Sundar; Sherchand, Jeevan Bahadur; Pokharel, Dinesh Binod; Pokhrel, Bharat Mani; Mishra, Shyam Kumar; Dhital, Subhash; Rijal, Basista

    2016-01-01

    Background. Superficial mycosis is a common fungal infection worldwide, mainly caused by dermatophytes. However, the prevalence of species varies geographically. In addition, fungal treatment is best guided according to species isolated. This study was carried out to determine the clinical as well as mycological profile of superficial mycoses in a tertiary care hospital, Nepal. Methods. This was a prospective case-control laboratory based study conducted over a period of six months from Janua...

  9. Emotional words can be embodied or disembodied: the role of superficial vs. deep types of processing

    OpenAIRE

    Abbassi, Ensie; Blanchette, Isabelle; Ansaldo, Ana I.; Ghassemzadeh, Habib; Joanette, Yves

    2015-01-01

    Emotional words are processed rapidly and automatically in the left hemisphere (LH) and slowly, with the involvement of attention, in the right hemisphere (RH). This review aims to find the reason for this difference and suggests that emotional words can be processed superficially or deeply due to the involvement of the linguistic and imagery systems, respectively. During superficial processing, emotional words likely make connections only with semantically associated words in the LH. This pa...

  10. Topographical anatomy of superficial veins, cutaneous nerves, and arteries at venipuncture sites in the cubital fossa.

    Science.gov (United States)

    Mikuni, Yuko; Chiba, Shoji; Tonosaki, Yoshikazu

    2013-01-01

    We investigated correlations among the superficial veins, cutaneous nerves, arteries, and venous valves in 128 cadaveric arms in order to choose safe venipuncture sites in the cubital fossa. The running patterns of the superficial veins were classified into four types (I-IV) and two subtypes (a and b). In types I and II, the median cubital vein (MCV) was connected obliquely between the cephalic and basilic veins in an N-shape, while the median antebrachial vein (MAV) opened into the MCV in type I and into the basilic vein in type II. In type III, the MCV did not exist. In type IV, additional superficial veins above the cephalic and basilic veins were developed around the cubital fossa. In types Ib-IVb, the accessory cephalic vein was developed under the same conditions as seen in types Ia-IVa, respectively. The lateral cutaneous nerve of the forearm descended deeply along the cephalic vein in 124 cases (97 %), while the medial cutaneous nerve of the forearm descended superficially along the basilic vein in 94 (73 %). A superficial brachial artery was found in 27 cases (21 %) and passed deeply under the ulnar side of the MCV. A median superficial antebrachial artery was found in 1 case (1 %), which passed deeply under the ulnar side of the MCV and ran along the MAV. Venous valves were found at 239 points in 28 cases with superficial veins, with a single valve seen at 79 points (33 %) and double valves at 160 points (67 %). At the time of intravenous injection, caution is needed regarding the locations of cutaneous nerves, brachial and superficial brachial arteries, and venous valves. The area ranging from the middle segment of the MCV to the confluence between the MCV and cephalic vein appears to be a relatively safe venipuncture site.

  11. Superficial siderosis of the central nervous system due to brachial plexus injury: a case report

    International Nuclear Information System (INIS)

    Setogutti, Enio Tadashi; Cassuriaga, Jefferson; Valduga, Simone Gianella; Lorenzzoni, Pablo Longhi; Severgnini, Giancarlo Muraro; Feldman, Carlos Jader

    2005-01-01

    Superficial siderosis can be caused by hemosiderin deposition o the leptomeninges and subpial layers of the neuro-axis due to recurrent subarachnoid haemorrhage. Probable intrathecal bleeding sites must be investigated. In ut t 50% of the patients the bleeding source may be identified and the progression of the disease can be interrupted. In this study, the authors present a case of superficial siderosis of the central nervous system developed two decades after a traumatic lesion of the brachial plexus.(author)

  12. Coexistence of adult-onset actinic prurigo and shampoo dermatitis: A case report

    Directory of Open Access Journals (Sweden)

    Tsung-Ju Lee

    2018-06-01

    Full Text Available Actinic prurigo is a rare and acquired idiopathic photodermatosis. It usually shows childhood onset and female predominance. Here, we present an unusual case of a male patient with coexistence of adult-onset actinic prurigo and shampoo-induced allergic contact dermatitis. He was initially diagnosed with actinic prurigo. However, after detailed examination of the distribution of the rash, careful collection of his history, and interpretation of the results of histopathologic analysis, photo test, patch test, and photopatch test, coexistence of adult-onset actinic prurigo and shampoo-induced allergic contact dermatitis associated with cocamidopropyl betaine was diagnosed. The rash improved after appropriate use of sunscreen and avoidance of shampoo containing this allergen. Dermatologists should be aware of the possibility of concurrent photodermatitis and contact dermatitis. Keywords: Actinic prurigo, Cocamidopropyl betaine, Contact dermatitis, Photosensitivity, Shampoo dermatitis

  13. Engineering an artificial amoeba propelled by nanoparticle-triggered actin polymerization

    Energy Technology Data Exchange (ETDEWEB)

    Yi Jinsoo; Schmidt, Jacob; Chien Aichi; Montemagno, Carlo D [Department of Bioengineering, University of California Los Angeles, 420 Westwood Plaza, 7523 Boelter Hall, Los Angeles, CA 90095-1600 (United States)], E-mail: montemcd@ucmail.uc.edu

    2009-02-25

    We have engineered an amoeba system combining nanofabricated inorganic materials with biological components, capable of propelling itself via actin polymerization. The nanofabricated materials have a mechanism similar to the locomotion of the Listeria monocytogenes, food poisoning bacteria. The propulsive force generation utilizes nanoparticles made from nickel and gold functionalized with the Listeria monocytogenes transmembrane protein, ActA. These Listeria-mimic nanoparticles were in concert with actin, actin binding proteins, ATP (adenosine triphosphate) and encapsulated within a lipid vesicle. This system is an artificial cell, such as a vesicle, where artificial nanobacteria and actin polymerization machinery are used in driving force generators inside the cell. The assembled structure was observed to crawl on a glass surface analogously to an amoeba, with the speed of the movement dependent on the amount of actin monomers and ATP present.

  14. Mutations in actin used for structural studies partially disrupt β-thymosin/WH2 domains interaction.

    Science.gov (United States)

    Deville, Célia; Girard-Blanc, Christine; Assrir, Nadine; Nhiri, Naïma; Jacquet, Eric; Bontems, François; Renault, Louis; Petres, Stéphane; van Heijenoort, Carine

    2016-10-01

    Understanding the structural basis of actin cytoskeleton remodeling requires stabilization of actin monomers, oligomers, and filaments in complex with partner proteins, using various biochemical strategies. Here, we report a dramatic destabilization of the dynamic interaction with a model β-thymosin/WH2 domain induced by mutations in actin. This result underlines that mutant actins should be used with prudence to characterize interactions with intrinsically disordered partners as destabilization of dynamic interactions, although identifiable by NMR, may be invisible to other structural techniques. It also highlights how both β-thymosin/WH2 domains and actin tune local structure and dynamics in regulatory processes involving intrinsically disordered domains. © 2016 Federation of European Biochemical Societies.

  15. A Nanodiamond-peptide Bioconjugate for Fluorescence and ODMR Microscopy of a Single Actin Filament.

    Science.gov (United States)

    Genjo, Takuya; Sotoma, Shingo; Tanabe, Ryotaro; Igarashi, Ryuji; Shirakawa, Masahiro

    2016-01-01

    Recently, the importance of conformational changes in actin filaments induced by mechanical stimulation of a cell has been increasingly recognized, especially in terms of mechanobiology. Despite its fundamental importance, however, long-term observation of a single actin filament by fluorescent microscopy has been difficult because of the low photostability of traditional fluorescent molecules. This paper reports a novel molecular labeling system for actin filaments using fluorescent nanodiamond (ND) particles harboring nitrogen-vacancy centers; ND has flexible chemical modifiability, extremely high photostability and biocompatibility, and provides a variety of physical information quantitatively via optically detected magnetic resonance (ODMR) measurements. We performed the chemical surface modification of an ND with the actin filament-specific binding peptide Lifeact and observed colocalization of pure Lifeact-modified ND and actin filaments by the ODMR selective imaging protocol, suggesting the capability of long-term observation and quantitative analysis of a single molecule by using an ND particle.

  16. Time-sequential observation of spindle and phragmoplast orientation in BY-2 cells with altered cortical actin microfilament patterning.

    Science.gov (United States)

    Kojo, Kei H; Yasuhara, Hiroki; Hasezawa, Seiichiro

    2014-01-01

    Precise division plane determination is essential for plant development. At metaphase, a dense actin microfilament meshwork appears on both sides of the cell center, forming a characteristic cortical actin microfilament twin peak pattern in BY-2 cells. We previously reported a strong correlation between altered cortical actin microfilament patterning and an oblique mitotic spindle orientation, implying that these actin microfilament twin peaks play a role in the regulation of mitotic spindle orientation. In the present study, time-sequential observation was used to reveal the progression from oblique phragmoplast to oblique cell plate orientation in cells with altered cortical actin microfilament patterning. In contrast to cells with normal actin microfilament twin peaks, oblique phragmoplast reorientation was rarely observed in cells with altered cortical actin microfilament patterning. These results support the important roles of cortical actin microfilament patterning in division plane orientation.

  17. The effect of membrane-regulated actin polymerization on a two-phase flow model for cell motility

    KAUST Repository

    Kimpton, L. S.; Whiteley, J. P.; Waters, S. L.; Oliver, J. M.

    2014-01-01

    travelling-wave solutions with biologically plausible actin network profiles in two simple models that enforce polymerization or depolymerization of the actin network at the ends of the travelling, 1D strip of cytoplasm. © 2014 The authors 2014. Published

  18. Vascular patterns of upper limb: an anatomical study with accent on superficial brachial artery

    Directory of Open Access Journals (Sweden)

    David Kachlik

    2011-02-01

    Full Text Available The aim of the study was to evaluate the terminal segmentation of the axillary artery and to present four cases of anomalous branching of the axillary artery, the superficial brachial artery (arteria brachialis superficialis, which is defined as the brachial artery that runs superficially to the median nerve. Totally, 130 cadaveric upper arms embalmed by classical formaldehyde technique from collections of the Department of Anatomy, Third Faculty of Medicine, Charles University in Prague, were macroscopically dissected with special focus on the branching arrangement of the axillary artery. The most distal part of the axillary artery (infrapectoral part terminated in four cases as a bifurcation into two terminal branches: the superficial brachial artery and profunda brachii artery, denominated according to their relation to the median nerve. The profunda brachii artery primarily gave rise to the main branches of the infrapectoral part of the axillary artery. The superficial brachial artery descended to the cubital fossa where it assumed the usual course of the brachial artery in two cases and in the other two cases its branches (the radial and ulnar arteries passed superficially to the flexors. The incidence of the superficial brachial artery in our study was 5% of cases. The reported incidence is a bit contradictory, from 0.12% to 25% of cases. The anatomical knowledge of the axillary region is of crucial importance for neurosurgeons and specialists using the radiodiagnostic techniques, particularly in cases involving traumatic injuries. The improved knowledge would allow more accurate diagnostic interpretations and surgical treatment.

  19. A Gly65Val substitution in an actin, GhACT_LI1, disrupts cell polarity and membrane anchoring of F-actin resulting in dwarf, lintless Li1 cotton plants

    Science.gov (United States)

    Actin polymerizes to form the cytoskeleton and organize polar growth in all eukaryotic cells. Species with numerous actin genes are especially useful for the dissection of actin molecular function due to redundancy and neofunctionalization. Here, we investigated the role of a cotton (Gossypium hi...

  20. Activation of the skeletal alpha-actin promoter during muscle regeneration.

    Science.gov (United States)

    Marsh, D R; Carson, J A; Stewart, L N; Booth, F W

    1998-11-01

    Little is known concerning promoter regulation of genes in regenerating skeletal muscles. In young rats, recovery of muscle mass and protein content is complete within 21 days. During the initial 5-10 days of regeneration, mRNA abundance for IGF-I, myogenin and MyoD have been shown to be dramatically increased. The skeletal alpha-actin promoter contains E box and serum response element (SRE) regulatory regions which are directly or indirectly activated by myogenin (or MyoD) and IGF-I proteins, respectively. We hypothesized that the skeletal alpha-actin promoter activity would increase during muscle regeneration, and that this induction would occur before muscle protein content returned to normal. Total protein content and the percentage content of skeletal alpha-actin protein was diminished at 4 and 8 days and re-accumulation had largely occurred by 16 days post-bupivacaine injection. Skeletal alpha-actin mRNA per whole muscle was decreased at day 8, and thereafter returned to control values. During regeneration at day 8, luciferase activity (a reporter of promoter activity) directed by -424 skeletal alpha-actin and -99 skeletal alpha-actin promoter constructs was increased by 700% and 250% respectively; however, at day 16, skeletal alpha-actin promoter activities were similar to control values. Thus, initial activation of the skeletal alpha-actin promoter is associated with regeneration of skeletal muscle, despite not being sustained during the later stages of regrowth. The proximal SRE of the skeletal alpha-actin promoter was not sufficient to confer a regeneration-induced promoter activation, despite increased serum response factor protein binding to this regulatory element in electrophoretic mobility shift assays. Skeletal alpha-actin promoter induction during regeneration is due to a combination of regulatory elements, at least including the SRE and E box.

  1. Regulation of myosin IIA and filamentous actin during insulin-stimulated glucose uptake in 3T3-L1 adipocytes

    International Nuclear Information System (INIS)

    Stall, Richard; Ramos, Joseph; Kent Fulcher, F.; Patel, Yashomati M.

    2014-01-01

    Insulin stimulated glucose uptake requires the colocalization of myosin IIA (MyoIIA) and the insulin-responsive glucose transporter 4 (GLUT4) at the plasma membrane for proper GLUT4 fusion. MyoIIA facilitates filamentous actin (F-actin) reorganization in various cell types. In adipocytes F-actin reorganization is required for insulin-stimulated glucose uptake. What is not known is whether MyoIIA interacts with F-actin to regulate insulin-induced GLUT4 fusion at the plasma membrane. To elucidate the relationship between MyoIIA and F-actin, we examined the colocalization of MyoIIA and F-actin at the plasma membrane upon insulin stimulation as well as the regulation of this interaction. Our findings demonstrated that MyoIIA and F-actin colocalized at the site of GLUT4 fusion with the plasma membrane upon insulin stimulation. Furthermore, inhibition of MyoII with blebbistatin impaired F-actin localization at the plasma membrane. Next we examined the regulatory role of calcium in MyoIIA-F-actin colocalization. Reduced calcium or calmodulin levels decreased colocalization of MyoIIA and F-actin at the plasma membrane. While calcium alone can translocate MyoIIA it did not stimulate F-actin accumulation at the plasma membrane. Taken together, we established that while MyoIIA activity is required for F-actin localization at the plasma membrane, it alone is insufficient to localize F-actin to the plasma membrane. - Highlights: • Insulin induces colocalization of MyoIIA and F-actin at the cortex in adipocytes. • MyoIIA is necessary but not sufficient to localize F-actin at the cell cortex. • MyoIIA-F-actin colocalization is regulated by calcium and calmodulin

  2. Quantitative studies of subdiffusion in living cells and actin networks

    DEFF Research Database (Denmark)

    Munteanu, Emilia-Laura; Olsen, Anja Lea; Tolic-Nørrelykke, Iva Marija

    2006-01-01

    Optical tweezers are a versatile tool in biophysics and have matured from a tool of manipulation to a tool of precise measurements. We argue here that the data analysis with advantage can be developed to a level of sophistication that matches that of the instrument. We review methods of analysis...... of optical tweezers data, primarily baed on the power spectra of time series of postions for trapped spherical objects. The majority of precise studies in the literature are performed on in vitro systems, whereas in the present work, an example of an in vivo system is presented for which precise power...... spectral analysis is both useful and necessary. The biological system is the cytoplasm of fission yeast, S. pombe, in which we observe subdiffusion of lipid granuli. in a search for the cause of subdiffusion, we chemically disrupt the actin network in the cytoplasm and further consider in vitro networks...

  3. Noisy Oscillations in the Actin Cytoskeleton of Chemotactic Amoeba

    Science.gov (United States)

    Negrete, Jose; Pumir, Alain; Hsu, Hsin-Fang; Westendorf, Christian; Tarantola, Marco; Beta, Carsten; Bodenschatz, Eberhard

    2016-09-01

    Biological systems with their complex biochemical networks are known to be intrinsically noisy. Here we investigate the dynamics of actin polymerization of amoeboid cells, which are close to the onset of oscillations. We show that the large phenotypic variability in the polymerization dynamics can be accurately captured by a generic nonlinear oscillator model in the presence of noise. We determine the relative role of the noise with a single dimensionless, experimentally accessible parameter, thus providing a quantitative description of the variability in a population of cells. Our approach, which rests on a generic description of a system close to a Hopf bifurcation and includes the effect of noise, can characterize the dynamics of a large class of noisy systems close to an oscillatory instability.

  4. Light emitting fabric for photodynamic treatment of actinic keratosis

    Science.gov (United States)

    Thecua, E.; Vicentini, C.; Vignion, A.-S.; Lecomte, F.; Deleporte, P.; Mortier, L.; Szeimies, R.-M.; Mordon, S.

    2017-02-01

    The integration of optical fibers into flexible textile structures, by using knitting or weaving processes can allow the development of flexible light sources. The paper aims to present a new technology: Light Emitting Fabrics (LEF), which can be used for example for PDT of Actinic Keratosis in Dermatology. The predetermined macro-bending of optical fibers, led to a homogeneous side emission of light over the entire surface of the fabric. Tests showed that additional curvatures when applying the LEF on non-planar surfaces had no impact on light delivery and proved that LEF can adapt to the human morphology. The ability of the LEF, coupled with a 635nm LASER source, to deliver a homogeneous light to lesions is currently assessed in a clinical trial for the treatment of AK of the scalp by PDT. The low irradiance and progressive activation of the photosensitizer ensure a pain reduction, compared to discomfort levels experienced by patients during a conventional PDT session.

  5. Actinic inspection of EUV reticles with arbitrary pattern design

    Science.gov (United States)

    Mochi, Iacopo; Helfenstein, Patrick; Rajeev, Rajendran; Fernandez, Sara; Kazazis, Dimitrios; Yoshitake, Shusuke; Ekinci, Yasin

    2017-10-01

    The re ective-mode EUV mask scanning lensless imaging microscope (RESCAN) is being developed to provide actinic mask inspection capabilities for defects and patterns with high resolution and high throughput, for 7 nm node and beyond. Here we, will report on our progress and present the results on programmed defect detection on random, logic-like patterns. The defects we investigated range from 200 nm to 50 nm size on the mask. We demonstrated the ability of RESCAN to detect these defects in die-to-die and die-to-database mode with a high signal to noise ratio. We also describe future plans for the upgrades to increase the resolution, the sensitivity, and the inspection speed of the demo tool.

  6. Region-Specific Involvement of Actin Rearrangement-Related Synaptic Structure Alterations in Conditioned Taste Aversion Memory

    Science.gov (United States)

    Bi, Ai-Ling; Wang, Yue; Li, Bo-Qin; Wang, Qian-Qian; Ma, Ling; Yu, Hui; Zhao, Ling; Chen, Zhe-Yu

    2010-01-01

    Actin rearrangement plays an essential role in learning and memory; however, the spatial and temporal regulation of actin dynamics in different phases of associative memory has not been fully understood. Here, using the conditioned taste aversion (CTA) paradigm, we investigated the region-specific involvement of actin rearrangement-related…

  7. The atypical Rho GTPase RhoD is a regulator of actin cytoskeleton dynamics and directed cell migration

    Energy Technology Data Exchange (ETDEWEB)

    Blom, Magdalena; Reis, Katarina [Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, SE-171 77 Stockholm (Sweden); Heldin, Johan [Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala SE-751 22 Uppsala (Sweden); Kreuger, Johan [Department of Medical Cell Biology, Science for Life Laboratory, Uppsala University, SE-751 23 Uppsala (Sweden); Aspenström, Pontus, E-mail: pontus.aspenstrom@ki.se [Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, SE-171 77 Stockholm (Sweden)

    2017-03-15

    RhoD belongs to the Rho GTPases, a protein family responsible for the regulation and organization of the actin cytoskeleton, and, consequently, many cellular processes like cell migration, cell division and vesicle trafficking. Here, we demonstrate that the actin cytoskeleton is dynamically regulated by increased or decreased protein levels of RhoD. Ectopic expression of RhoD has previously been shown to give an intertwined weave of actin filaments. We show that this RhoD-dependent effect is detected in several cell types and results in a less dynamic actin filament system. In contrast, RhoD depletion leads to increased actin filament-containing structures, such as cortical actin, stress fibers and edge ruffles. Moreover, vital cellular functions such as cell migration and proliferation are defective when RhoD is silenced. Taken together, we present data suggesting that RhoD is an important component in the control of actin dynamics and directed cell migration. - Highlights: • Increased RhoD expression leads to loss of actin structures, e.g. stress fibers and gives rise to decreased actin dynamics. • RhoD knockdown induces various actin-containing structures such as edge ruffles, stress fibers and cortical actin, in a cell-type specific manner. • RhoD induces specific actin rearrangements depending on its subcellular localization. • RhoD knockdown has effects on cellular processes, such as directed cell migration and proliferation.

  8. Capu and Spire Assemble a Cytoplasmic Actin~Mesh that Maintains Microtubule Organization in the Drosophila Oocyte

    DEFF Research Database (Denmark)

    Dahlgaard, K.; Raposo, A.A.S.F.; Niccoli, T.

    2007-01-01

    Mutants in the actin nucleators Cappuccino and Spire disrupt the polarized microtubule network in the Drosophila oocyte that defines the anterior-posterior axis, suggesting that microtubule organization depends on actin. Here, we show that Cappuccino and Spire organize an isotropic mesh of actin...

  9. Dissecting the actin cortex density and membrane-cortex distance in living cells by super-resolution microscopy

    DEFF Research Database (Denmark)

    Clausen, M. P.; Colin-York, H.; Schneider, Falk

    2017-01-01

    and accurately measure the density distribution of the cortical actin cytoskeleton and the distance between the actin cortex and the membrane in live Jurkat T-cells. We found an asymmetric cortical actin density distribution with a mean width of 230 (+105/-125) nm. The spatial distances measured between...

  10. The atypical Rho GTPase RhoD is a regulator of actin cytoskeleton dynamics and directed cell migration

    International Nuclear Information System (INIS)

    Blom, Magdalena; Reis, Katarina; Heldin, Johan; Kreuger, Johan; Aspenström, Pontus

    2017-01-01

    RhoD belongs to the Rho GTPases, a protein family responsible for the regulation and organization of the actin cytoskeleton, and, consequently, many cellular processes like cell migration, cell division and vesicle trafficking. Here, we demonstrate that the actin cytoskeleton is dynamically regulated by increased or decreased protein levels of RhoD. Ectopic expression of RhoD has previously been shown to give an intertwined weave of actin filaments. We show that this RhoD-dependent effect is detected in several cell types and results in a less dynamic actin filament system. In contrast, RhoD depletion leads to increased actin filament-containing structures, such as cortical actin, stress fibers and edge ruffles. Moreover, vital cellular functions such as cell migration and proliferation are defective when RhoD is silenced. Taken together, we present data suggesting that RhoD is an important component in the control of actin dynamics and directed cell migration. - Highlights: • Increased RhoD expression leads to loss of actin structures, e.g. stress fibers and gives rise to decreased actin dynamics. • RhoD knockdown induces various actin-containing structures such as edge ruffles, stress fibers and cortical actin, in a cell-type specific manner. • RhoD induces specific actin rearrangements depending on its subcellular localization. • RhoD knockdown has effects on cellular processes, such as directed cell migration and proliferation.

  11. Calcium and actin in the saga of awakening oocytes

    Energy Technology Data Exchange (ETDEWEB)

    Santella, Luigia, E-mail: santella@szn.it; Limatola, Nunzia; Chun, Jong T.

    2015-04-24

    The interaction of the spermatozoon with the egg at fertilization remains one of the most fascinating mysteries of life. Much of our scientific knowledge on fertilization comes from studies on sea urchin and starfish, which provide plenty of gametes. Large and transparent, these eggs have served as excellent model systems for studying egg activation and embryo development in seawater, a plain natural medium. Starfish oocytes allow the study of the cortical, cytoplasmic and nuclear changes during the meiotic maturation process, which can also be triggered in vitro by hormonal stimulation. These morphological and biochemical changes ensure successful fertilization of the eggs at the first metaphase. On the other hand, sea urchin eggs are fertilized after the completion of meiosis, and are particularly suitable for the study of sperm–egg interaction, early events of egg activation, and embryonic development, as a large number of mature eggs can be fertilized synchronously. Starfish and sea urchin eggs undergo abrupt changes in the cytoskeleton and ion fluxes in response to the fertilizing spermatozoon. The plasma membrane and cortex of an egg thus represent “excitable media” that quickly respond to the stimulus with the Ca{sup 2+} swings and structural changes. In this article, we review some of the key findings on the rapid dynamic rearrangements of the actin cytoskeleton in the oocyte/egg cortex upon hormonal or sperm stimulation and their roles in the modulation of the Ca{sup 2+} signals and in the control of monospermic fertilization. - Highlights: • Besides microtubules, microfilaments may anchor the nucleus to oocyte surface. • The cortical Ca{sup 2+} flash and wave at fertilization mirror electrical membrane change. • Artificial egg activation lacks microvilli extension in the perivitelline space. • Calcium is necessary but not sufficient for cortical granules exocytosis. • Actin cytoskeleton modulates Ca{sup 2+} release at oocyte maturation

  12. Calcium and actin in the saga of awakening oocytes

    International Nuclear Information System (INIS)

    Santella, Luigia; Limatola, Nunzia; Chun, Jong T.

    2015-01-01

    The interaction of the spermatozoon with the egg at fertilization remains one of the most fascinating mysteries of life. Much of our scientific knowledge on fertilization comes from studies on sea urchin and starfish, which provide plenty of gametes. Large and transparent, these eggs have served as excellent model systems for studying egg activation and embryo development in seawater, a plain natural medium. Starfish oocytes allow the study of the cortical, cytoplasmic and nuclear changes during the meiotic maturation process, which can also be triggered in vitro by hormonal stimulation. These morphological and biochemical changes ensure successful fertilization of the eggs at the first metaphase. On the other hand, sea urchin eggs are fertilized after the completion of meiosis, and are particularly suitable for the study of sperm–egg interaction, early events of egg activation, and embryonic development, as a large number of mature eggs can be fertilized synchronously. Starfish and sea urchin eggs undergo abrupt changes in the cytoskeleton and ion fluxes in response to the fertilizing spermatozoon. The plasma membrane and cortex of an egg thus represent “excitable media” that quickly respond to the stimulus with the Ca 2+ swings and structural changes. In this article, we review some of the key findings on the rapid dynamic rearrangements of the actin cytoskeleton in the oocyte/egg cortex upon hormonal or sperm stimulation and their roles in the modulation of the Ca 2+ signals and in the control of monospermic fertilization. - Highlights: • Besides microtubules, microfilaments may anchor the nucleus to oocyte surface. • The cortical Ca 2+ flash and wave at fertilization mirror electrical membrane change. • Artificial egg activation lacks microvilli extension in the perivitelline space. • Calcium is necessary but not sufficient for cortical granules exocytosis. • Actin cytoskeleton modulates Ca 2+ release at oocyte maturation and fertilization

  13. Extending the molecular clutch beyond actin-based cell motility

    International Nuclear Information System (INIS)

    Havrylenko, Svitlana; Mezanges, Xavier; Batchelder, Ellen; Plastino, Julie

    2014-01-01

    Many cell movements occur via polymerization of the actin cytoskeleton beneath the plasma membrane at the front of the cell, forming a protrusion called a lamellipodium, while myosin contraction squeezes forward the back of the cell. In what is known as the ‘molecular clutch’ description of cell motility, forward movement results from the engagement of the acto-myosin motor with cell-matrix adhesions, thus transmitting force to the substrate and producing movement. However during cell translocation, clutch engagement is not perfect, and as a result, the cytoskeleton slips with respect to the substrate, undergoing backward (retrograde) flow in the direction of the cell body. Retrograde flow is therefore inversely proportional to cell speed and depends on adhesion and acto-myosin dynamics. Here we asked whether the molecular clutch was a general mechanism by measuring motility and retrograde flow for the Caenorhabditis elegans sperm cell in different adhesive conditions. These cells move by adhering to the substrate and emitting a dynamic lamellipodium, but the sperm cell does not contain an acto-myosin cytoskeleton. Instead the lamellipodium is formed by the assembly of major sperm protein, which has no biochemical or structural similarity to actin. We find that these cells display the same molecular clutch characteristics as acto-myosin containing cells. We further show that retrograde flow is produced both by cytoskeletal assembly and contractility in these cells. Overall this study shows that the molecular clutch hypothesis of how polymerization is transduced into motility via adhesions is a general description of cell movement regardless of the composition of the cytoskeleton. (paper)

  14. Evaluating optimal superficial limb perfusion at different angles using non-invasive micro-lightguide spectrophotometry.

    Science.gov (United States)

    Darmanin, Geraldine; Jaggard, Matthew; Hettiaratchy, Shehan; Nanchahal, Jagdeep; Jain, Abhilash

    2013-06-01

    It is common practice to elevate the limbs postoperatively to reduce oedema and hence optimise perfusion and facilitate rehabilitation. However, elevation may be counterproductive as it reduces the mean perfusion pressure. There are no clear data on the optimal position of the limbs even in normal subjects. The optimal position of limbs was investigated in 25 healthy subjects using a non-invasive micro-lightguide spectrophotometry system "O2C", which indirectly measures skin and superficial tissue perfusion through blood flow, oxygen saturation and relative haemoglobin concentration. We found a reduction in skin and superficial tissue blood flow of 17% (p=0.0001) on arm elevation (180° shoulder flexion) as compared to heart level and an increase in skin and superficial tissue blood flow of 25% (p=0.02) on forearm elevation of 45°. Lower limb skin and superficial tissue blood flow decreased by 15% (p=0.004) on elevation to 47 cm and by 70% on dependency (p=0.0001) compared to heart level. However, on elevation of the lower limb there was also a 28% reduction in superficial venous pooling (p=0.0001) compared to heart level. In the normal limb, the position for optimal superficial perfusion of the upper limb is with the arm placed at heart level and forearm at 45°. In the lower limb the optimal position for superficial perfusion would be at heart level. However, some degree of elevation may be useful if there is an element of venous congestion. Copyright © 2013 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.

  15. Computations in the deep vs superficial layers of the cerebral cortex.

    Science.gov (United States)

    Rolls, Edmund T; Mills, W Patrick C

    2017-11-01

    A fundamental question is how the cerebral neocortex operates functionally, computationally. The cerebral neocortex with its superficial and deep layers and highly developed recurrent collateral systems that provide a basis for memory-related processing might perform somewhat different computations in the superficial and deep layers. Here we take into account the quantitative connectivity within and between laminae. Using integrate-and-fire neuronal network simulations that incorporate this connectivity, we first show that attractor networks implemented in the deep layers that are activated by the superficial layers could be partly independent in that the deep layers might have a different time course, which might because of adaptation be more transient and useful for outputs from the neocortex. In contrast the superficial layers could implement more prolonged firing, useful for slow learning and for short-term memory. Second, we show that a different type of computation could in principle be performed in the superficial and deep layers, by showing that the superficial layers could operate as a discrete attractor network useful for categorisation and feeding information forward up a cortical hierarchy, whereas the deep layers could operate as a continuous attractor network useful for providing a spatially and temporally smooth output to output systems in the brain. A key advance is that we draw attention to the functions of the recurrent collateral connections between cortical pyramidal cells, often omitted in canonical models of the neocortex, and address principles of operation of the neocortex by which the superficial and deep layers might be specialized for different types of attractor-related memory functions implemented by the recurrent collaterals. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. The interplay between viscoelastic and thermodynamic properties determines the birefringence of F-actin gels.

    Science.gov (United States)

    Helfer, Emmanuèle; Panine, Pierre; Carlier, Marie-France; Davidson, Patrick

    2005-07-01

    F-actin gels of increasing concentrations (25-300 microM) display in vitro a progressive onset of birefringence due to orientational ordering of actin filaments. At F-actin concentrations mechanical stresses stored in the gels. In contrast, at F-actin concentrations > or =100 microM, gels display spontaneous birefringence recovery, at rest, which is the sign of true nematic ordering, in good agreement with statistical physics models of the isotropic/nematic transition. Well-aligned samples of F-actin gels could be produced and their small-angle x-ray scattering patterns are quite anisotropic. These patterns show no sign of filament positional short-range order and could be modeled by averaging the form factor with the Maier-Saupe nematic distribution function. The derived nematic order parameter S of the gels ranged from S = 0.7 at 300 microM to S = 0.4 at 25 microM. Both birefringence and small-angle x-ray scattering data indicate that, even in absence of cross-linking proteins, spontaneous cooperative alignment of actin filaments may arise in motile regions of living cells where F-actin concentrations can reach values of a few 100 microM.

  17. Interactions between globular proteins and F-actin in isotonic saline solution.

    Science.gov (United States)

    Lakatos, S; Minton, A P

    1991-10-05

    Solutions of each of three different globular proteins (cytochrome c, chromophorically labeled serum albumin, and chromophorically labeled aldolase), mixed with another unlabeled globular protein or with fibrous actin, were prepared in pH 8.0 Tris-HCl buffer containing 0.15 M NaCl. Each solution was centrifuged at low speed, at 5 degrees C, until unassociated globular protein in solution achieved sedimentation equilibrium. Individual absorbance gradients of both macrosolutes in the mixtures subsequent to centrifugation were obtained via optical scans of the centrifuge tubes at two wavelengths. The gradients of each macrosolute in mixtures of two globular proteins revealed no association of globular proteins under the conditions of these experiments, but perturbation of the gradients of serum albumin, aldolase, and cytochrome c in the presence of F-actin indicated association of all three globular proteins with F-actin. Perturbation of actin gradients in the presence of serum albumin and aldolase suggested partial depolymerization of the F-actin by the globular protein. Analysis of the data with a simple phenomenological model relating free globular protein, bound globular protein, and total actin concentration provided estimates of the respective equilibrium constants for association of serum albumin and aldolase with F-actin, under the conditions of these experiments, of the order of 0.1 microM-1.

  18. A glycolytic metabolon in Saccharomyces cerevisiae is stabilized by F-actin.

    Science.gov (United States)

    Araiza-Olivera, Daniela; Chiquete-Felix, Natalia; Rosas-Lemus, Mónica; Sampedro, José G; Peña, Antonio; Mujica, Adela; Uribe-Carvajal, Salvador

    2013-08-01

    In the Saccharomyces cerevisiae glycolytic pathway, 11 enzymes catalyze the stepwise conversion of glucose to two molecules of ethanol plus two CO₂ molecules. In the highly crowded cytoplasm, this pathway would be very inefficient if it were dependent on substrate/enzyme diffusion. Therefore, the existence of a multi-enzymatic glycolytic complex has been suggested. This complex probably uses the cytoskeleton to stabilize the interaction of the various enzymes. Here, the role of filamentous actin (F-actin) in stabilization of a putative glycolytic metabolon is reported. Experiments were performed in isolated enzyme/actin mixtures, cytoplasmic extracts and permeabilized yeast cells. Polymerization of actin was promoted using phalloidin or inhibited using cytochalasin D or latrunculin. The polymeric filamentous F-actin, but not the monomeric globular G-actin, stabilized both the interaction of isolated glycolytic pathway enzyme mixtures and the whole fermentation pathway, leading to higher fermentation activity. The associated complexes were resistant against inhibition as a result of viscosity (promoted by the disaccharide trehalose) or inactivation (using specific enzyme antibodies). In S. cerevisiae, a glycolytic metabolon appear to assemble in association with F-actin. In this complex, fermentation activity is enhanced and enzymes are partially protected against inhibition by trehalose or by antibodies. © 2013 FEBS.

  19. Actin Cytoskeleton Manipulation by Effector Proteins Secreted by Diarrheagenic Escherichia coli Pathotypes

    Directory of Open Access Journals (Sweden)

    Fernando Navarro-Garcia

    2013-01-01

    Full Text Available The actin cytoskeleton is a dynamic structure necessary for cell and tissue organization, including the maintenance of epithelial barriers. Disruption of the epithelial barrier coincides with alterations of the actin cytoskeleton in several disease states. These disruptions primarily affect the paracellular space, which is normally regulated by tight junctions. Thereby, the actin cytoskeleton is a common and recurring target of bacterial virulence factors. In order to manipulate the actin cytoskeleton, bacteria secrete and inject toxins and effectors to hijack the host cell machinery, which interferes with host-cell pathways and with a number of actin binding proteins. An interesting model to study actin manipulation by bacterial effectors is Escherichia coli since due to its genome plasticity it has acquired diverse genetic mobile elements, which allow having different E. coli varieties in one bacterial species. These E. coli pathotypes, including intracellular and extracellular bacteria, interact with epithelial cells, and their interactions depend on a specific combination of virulence factors. In this paper we focus on E. coli effectors that mimic host cell proteins to manipulate the actin cytoskeleton. The study of bacterial effector-cytoskeleton interaction will contribute not only to the comprehension of the molecular causes of infectious diseases but also to increase our knowledge of cell biology.

  20. Espins and the actin cytoskeleton of hair cell stereocilia and sensory cell microvilli

    Science.gov (United States)

    Sekerková, Gabriella; Zheng, Lili; Loomis, Patricia A.; Mugnaini, Enrico; Bartles, James R.

    2008-01-01

    The espins are novel actin-bundling proteins that are produced in multiple isoforms from a single gene. They are present at high concentration in the parallel actin bundle of hair cell stereocilia and are the target of deafness mutations in mice and humans. Espins are also enriched in the microvilli of taste receptor cells, solitary chemoreceptor cells, vomeronasal sensory neurons and Merkel cells, suggesting that espins play important roles in the microvillar projections of vertebrate sensory cells. Espins are potent actin-bundling proteins that are not inhibited by Ca2+. In cells, they efficiently elongate parallel actin bundles and, thereby, help determine the steady-state length of microvilli and stereocilia. Espins bind actin monomer via their WH2 domain and can assemble actin bundles in cells. Certain espin isoforms can also bind phosphatidylinositol 4,5-bisphosphate, profilins or SH3 proteins. These biological activities distinguish espins from other actin-bundling proteins and may make them well-suited to sensory cells. PMID:16909209

  1. Actin-based gravity-sensing mechanisms in unicellular plant model systems

    Science.gov (United States)

    Braun, Markus; Limbach, Christoph

    2005-08-01

    Considerable progress has been made in the understanding of the molecular and cellular mechanisms underlying gravity sensing and gravity-oriented polarized growth in single-celled rhizoids and protonemata of the characean algae. It is well known that the actin cytoskeleton plays a key role in these processes. Numerous actin-binding proteins control apical actin polymerization and the dynamic remodeling of the actin arrangement. An actomyosin-based system mediates the delivery and incorporation of secretory vesicles at the growing tip and coordinates the tip-high gradient of cytoplasmic free calcium which is required for local exocytosis. Additionally, the actomyosin system precisely controls the position of statoliths and, upon a change in orientation relative to the gravity vector, directs sedimenting statoliths to the confined graviperception sites of the plasma membrane where gravitropic signalling is initiated. The upward growth response of protonemata is preceded by an actin-dependent relocalization of the Ca2+-gradient to the upper flank. The downward growth response of rhizoids, however, is caused by differential growth of the opposite flankes due to a local reduction of cytoplasmic free calcium limited to the plasma membrane area where statoliths are sedimented. Thus, constant actin polymerization in the growing tip and the spatiotemporal control of actin remodeling are essential for gravity sensing and gravity-oriented polarized growth of characean rhizoids and protonemata.

  2. Localization of Myosin and Actin in the Pelage and Whisker Hair Follicles of Rat

    International Nuclear Information System (INIS)

    Morioka, Kiyokazu; Matsuzaki, Toshiyuki; Takata, Kuniaki

    2006-01-01

    The combined effects of myosin II and actin enable muscle and nonmuscle cells to generate forces required for muscle contraction, cell division, cell migration, cellular morphological changes, the maintenance of cellular tension and polarity, and so on. However, except for the case of muscle contraction, the details are poorly understood. We focus on nonmuscle myosin and actin in the formation and maintenance of hair and skin, which include highly active processes in mammalian life with respect to the cellular proliferation, differentiation, and movement. The localization of nonmuscle myosin II and actin in neonatal rat dorsal skin, mystacial pad, hair follicles, and vibrissal follicles was studied by immunohistochemical technique to provide the basis for the elucidation of the roles of these proteins. Specificities of the antibodies were verified by using samples from the relevant tissues and subjecting them to immunoblotting test prior to morphological analyses. The myosin and actin were abundant and colocalized in the spinous and granular layers but scarce in the basal layer of the dorsal and mystacial epidermis. In hair and vibrissal follicles, nonmuscle myosin and actin were colocalized in the outer root sheath and some hair matrix cells adjoining dermal papillae. In contrast, most areas of the inner root sheath and hair matrix appeared to comprise very small amounts of myosin and actin. Hair shaft may comprise significant myosin during the course of its keratinization. These results suggest that the actin-myosin system plays a part in cell movement, differentiation, protection and other key functions of skin and hair cells

  3. Live cell imaging of mitochondrial movement along actin cables in budding yeast.

    Science.gov (United States)

    Fehrenbacher, Kammy L; Yang, Hyeong-Cheol; Gay, Anna Card; Huckaba, Thomas M; Pon, Liza A

    2004-11-23

    Mitochondrial inheritance is essential for cell division. In budding yeast, mitochondrial movement from mother to daughter requires (1) actin cables, F-actin bundles that undergo retrograde movement during elongation from buds into mother cells; (2) the mitochore, a mitochondrial protein complex implicated in linking mitochondria to actin cables; and (3) Arp2/3 complex-mediated force generation on mitochondria. We observed three new classes of mitochondrial motility: anterograde movement at velocities of 0.2-0.33 microm/s, retrograde movement at velocities of 0.26-0.51 microm/s, and no net anterograde or retrograde movement. In all cases, motile mitochondria were associated with actin cables undergoing retrograde flow at velocities of 0.18-0.62 microm/s. Destabilization of actin cables or mutations of the mitochore blocked all mitochondrial movements. In contrast, mutations in the Arp2/3 complex affected anterograde but not retrograde mitochondrial movements. Actin cables are required for movement of mitochondria, secretory vesicles, mRNA, and spindle alignment elements in yeast. We provide the first direct evidence that one of the proposed cargos use actin cables as tracks. In the case of mitochondrial inheritance, anterograde movement drives transfer of the organelle from mothers to buds, while retrograde movement contributes to retention of the organelle in mother cells. Interaction of mitochondria with actin cables is required for anterograde and retrograde movement. In contrast, force generation on mitochondria is required only for anterograde movement. Finally, we propose a novel mechanism in which actin cables serve as "conveyor belts" that drive retrograde organelle movement.

  4. Comparative genome analysis reveals a conserved family of actin-like proteins in apicomplexan parasites

    Directory of Open Access Journals (Sweden)

    Sibley L David

    2005-12-01

    Full Text Available Abstract Background The phylum Apicomplexa is an early-branching eukaryotic lineage that contains a number of important human and animal pathogens. Their complex life cycles and unique cytoskeletal features distinguish them from other model eukaryotes. Apicomplexans rely on actin-based motility for cell invasion, yet the regulation of this system remains largely unknown. Consequently, we focused our efforts on identifying actin-related proteins in the recently completed genomes of Toxoplasma gondii, Plasmodium spp., Cryptosporidium spp., and Theileria spp. Results Comparative genomic and phylogenetic studies of apicomplexan genomes reveals that most contain only a single conventional actin and yet they each have 8–10 additional actin-related proteins. Among these are a highly conserved Arp1 protein (likely part of a conserved dynactin complex, and Arp4 and Arp6 homologues (subunits of the chromatin-remodeling machinery. In contrast, apicomplexans lack canonical Arp2 or Arp3 proteins, suggesting they lost the Arp2/3 actin polymerization complex on their evolutionary path towards intracellular parasitism. Seven of these actin-like proteins (ALPs are novel to apicomplexans. They show no phylogenetic associations to the known Arp groups and likely serve functions specific to this important group of intracellular parasites. Conclusion The large diversity of actin-like proteins in apicomplexans suggests that the actin protein family has diverged to fulfill various roles in the unique biology of intracellular parasites. Conserved Arps likely participate in vesicular transport and gene expression, while apicomplexan-specific ALPs may control unique biological traits such as actin-based gliding motility.

  5. Superficial corneal foreign body: laboratory and epidemiologic aspects Corpo estranho corneal superficial: aspectos laboratoriais e epidemiológicos

    Directory of Open Access Journals (Sweden)

    Ednajar Tavares Macedo Filho

    2005-12-01

    Full Text Available PURPOSE: To determine by bacterioscopy and culture the microorganisms carried by corneal foreign body and their sensitivity to antibiotics by antibiotic sensitivity test. METHODS: A prospective study was carried out and information was collected on 101 patients who presented with corneal foreign body at the São Paulo Hospital Eye Emergency Service. Prior to any treatment, a sample of the ipsilateral inferior conjunctival fornix and the foreign body were collected and immersed in thioglycolate broth. Samples were sown on solid culture media including blood, chocolate and Sabouraud agar. Bacterioscopic examination using Gram and Giemsa staining and sensitivity test were performed. Positive foreign body culture results were compared to ipsilateral conjunctival fornix culture to exclude possible normal flora growth. RESULTS: Approximately 92% of patients were males with a mean age of 35 years and in 62.4% (95% confidence interval: 52.2 - 71.8% had the right eye was affected. Foreign body positive cultures were achieved in 32.7% (95% confidence interval: 23.7 - 42.7% of the cases. The microorganisms isolated from the foreign body culture were identified as: Streptococcus, alpha-hemolytic (n=4, Staphylococcus aureus (n=4, Staphylococcus, coagulase-negative (n=4, Corynebacterium xerosis (n=3, unidentified Gram-positive bacillus (n=2, Moraxella sp (n=1, Serratia sp (n=1, Acynetobacter sp (n=1. The microbial sensitivity test showed that 95% of the cases were sensitive to chloramphenicol and 90% were sensitive to ciprofloxacin, these being antibiotics used in our routine service. CONCLUSION: Superficial corneal foreign body acts as important contaminant vector and the great majority of isolated bacteria were sensitive to the antibiotic prophylaxis used in the treatment.OBJETIVOS: Identificar por meio de cultura e bacterioscopia os microrganismos veiculados por corpo estranho corneal e sua sensibilidade aos antibióticos em teste de antibiograma. M

  6. Structural Polymorphism of the Actin-Espin System: A Prototypical System of Filaments and Linkers in Stereocilia

    International Nuclear Information System (INIS)

    Purdy, Kirstin R.; Wong, Gerard C. L.; Bartles, James R.

    2007-01-01

    We examine the interaction between cytoskeletal F-actin and espin 3A, a prototypical actin bundling protein found in sensory cell microvilli, including ear cell stereocilia. Espin induces twist distortions in F-actin as well as facilitates bundle formation. Mutations in one of the two F-actin binding sites of espin, which have been implicated in deafness, can tune espin-actin interactions and radically transform the system's phase behavior. These results are compared to recent theoretical work on the general phase behavior linker-rod systems

  7. Self-assembly of actin monomers into long filaments: Brownian Dynamics simulations

    DEFF Research Database (Denmark)

    Shillcock, Julian C.

    2009-01-01

    Brownian dynamics simulations are used to study the dynamical process of self-assembly of actin monomers into long filaments containing up to 1000 actin protomers. In order to overcome the large separation of time scales between the diffusive motion of the freemonomers and the relatively slow....../detachment events. When a single filament is allowed to grow in a bath of constant concentration of free ADP-actin monomers, its growth rate increases linearly with the free monomer concentration in quantitative agreement with in vitro experiments. Theresults also show that the waiting time is governed by...

  8. Clinicopahological features of superficial basaloid squamous cell carcinoma of the esophagus.

    Science.gov (United States)

    Oguma, J; Ozawa, S; Kazuno, A; Nitta, M; Ninomiya, Y; Tomita, S

    2017-12-01

    Basaloid squamous cell carcinoma (BSC) of the esophagus is classified as an epithelial malignant tumor and is a rare variant of squamous cell carcinoma (SCC). Most previous reports have suggested that advanced BSC has a poorer prognosis than typical SCC because of its high biological malignancy, but the biological activity of superficial BSC remains unclear. Twenty cases of superficial BSC, which underwent surgical resection in Tokai University Hospital between January 2004 and December 2013, were analyzed retrospectively. Among these cases, 19 cases with a T1 depth of invasion (BSC group) were compared with 180 cases of SCC that were resected during the same period and were pathologically diagnosed as T1 (SCC group). The frequency of lymph node metastasis in the T1 BSC group was significantly lower (2 patients, 11%) than that in the SCC group (84 patients, 47%) (P = 0.005). The frequency of lymphatic invasion in the BSC group was also lower (9 patients, 47%) than that in the SCC group (131 patients, 73%) (P = 0.021). The pathological type of the metastatic lymph node was BSC in all the superficial BSC cases with lymph node metastasis. This study demonstrated that lymph node metastasis was less likely to occur in cases with superficial BSC than in cases with superficial SCC. © The Authors 2017. Published by Oxford University Press on behalf of International Society for Diseases of the Esophagus. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  9. Clinicomycological Characterization of Superficial Mycoses from a Tertiary Care Hospital in Nepal.

    Science.gov (United States)

    Khadka, Sundar; Sherchand, Jeevan Bahadur; Pokharel, Dinesh Binod; Pokhrel, Bharat Mani; Mishra, Shyam Kumar; Dhital, Subhash; Rijal, Basista

    2016-01-01

    Background . Superficial mycosis is a common fungal infection worldwide, mainly caused by dermatophytes. However, the prevalence of species varies geographically. In addition, fungal treatment is best guided according to species isolated. This study was carried out to determine the clinical as well as mycological profile of superficial mycoses in a tertiary care hospital, Nepal. Methods . This was a prospective case-control laboratory based study conducted over a period of six months from January to June 2014 at Tribhuvan University Teaching Hospital, Nepal. A total of 200 specimens were collected from the patients suspected of superficial mycoses. The specimens were macroscopically as well as microscopically examined. The growth was observed up to 4 weeks. Results. Out of total 200 specimens from the patients suspected of superficial mycoses, tinea corporis 50 (25%) was most common clinical types. KOH mount was positive in 89 (44.5%) and culture was positive in 111 (55.5%). Trichophyton mentagrophytes 44 (39.6%) was the most common isolate. Conclusions. The diagnostic yields of KOH mount and culture were found to be complementary to each other. Thus both the methods added with clinical findings are equally important to establish superficial mycosis.

  10. Clinicomycological Characterization of Superficial Mycoses from a Tertiary Care Hospital in Nepal

    Directory of Open Access Journals (Sweden)

    Sundar Khadka

    2016-01-01

    Full Text Available Background. Superficial mycosis is a common fungal infection worldwide, mainly caused by dermatophytes. However, the prevalence of species varies geographically. In addition, fungal treatment is best guided according to species isolated. This study was carried out to determine the clinical as well as mycological profile of superficial mycoses in a tertiary care hospital, Nepal. Methods. This was a prospective case-control laboratory based study conducted over a period of six months from January to June 2014 at Tribhuvan University Teaching Hospital, Nepal. A total of 200 specimens were collected from the patients suspected of superficial mycoses. The specimens were macroscopically as well as microscopically examined. The growth was observed up to 4 weeks. Results. Out of total 200 specimens from the patients suspected of superficial mycoses, tinea corporis 50 (25% was most common clinical types. KOH mount was positive in 89 (44.5% and culture was positive in 111 (55.5%. Trichophyton mentagrophytes 44 (39.6% was the most common isolate. Conclusions. The diagnostic yields of KOH mount and culture were found to be complementary to each other. Thus both the methods added with clinical findings are equally important to establish superficial mycosis.

  11. Gross anatomy of superficial fascia and future localised fat deposit areas of the abdomen in foetus

    Directory of Open Access Journals (Sweden)

    Pramod Kumar

    2013-01-01

    Full Text Available Background: The development and popularity of body contouring procedures such as liposuction and abdominoplasty has renewed interest in the anatomy of the superficial fascia and subcutaneous fat deposits of the abdomen. The study of anatomy of fascia and fetal adipose tissue was proposed as it may be of value in understanding the possible programing of prevention of obesity. Objectives: The present study was undertaken to understand the gross anatomy of superficial fascia of abdomen and to study the gross anatomy of future localized fat deposits (LFDs area of abdomen in fetus. Materials and Methods: Four fetus (two male & two female of four month of intrauterine life were dissected. Attachments & layers of superficial fascia and future subcutaneous fat deposit area of upper and lower abdomen were noted. Results: Superficial fascia of the abdomen was multi layered in mid line and number of layers reduced laterally as in adult. The future abdominal LFD (localized fat deposits area in fetus shows brownish-white blubbary tissue without well-defined adult fat lobules. Conclusion: The attachment and gross anatomy of superficial fascia of the fetus was similar to that in adults. The future LFD areas showed brownish white blubbary tissue with ill-defined fat lobules.

  12. F-actin distribution at nodes of Ranvier and Schmidt-Lanterman incisures in mammalian sciatic nerves.

    Science.gov (United States)

    Kun, Alejandra; Canclini, Lucía; Rosso, Gonzalo; Bresque, Mariana; Romeo, Carlos; Hanusz, Alicia; Cal, Karina; Calliari, Aldo; Sotelo Silveira, José; Sotelo, José R

    2012-07-01

    Very little is known about the function of the F-actin cytoskeleton in the regeneration and pathology of peripheral nerve fibers. The actin cytoskeleton has been associated with maintenance of tissue structure, transmission of traction and contraction forces, and an involvement in cell motility. Therefore, the state of the actin cytoskeleton strongly influences the mechanical properties of cells and intracellular transport therein. In this work, we analyze the distribution of F-actin at Schmidt-Lanterman Incisures (SLI) and nodes of Ranvier (NR) domains in normal, regenerating and pathologic Trembler J (TrJ/+) sciatic nerve fibers, of rats and mice. F-actin was quantified and it was found increased in TrJ/+, both in SLI and NR. However, SLI and NR of regenerating rat sciatic nerve did not show significant differences in F-actin, as compared with normal nerves. Cytochalasin-D and Latrunculin-A were used to disrupt the F-actin network in normal and regenerating rat sciatic nerve fibers. Both drugs disrupt F-actin, but in different ways. Cytochalasin-D did not disrupt Schwann cell (SC) F-actin at the NR. Latrunculin-A did not disrupt F-actin at the boundary region between SC and axon at the NR domain. We surmise that the rearrangement of F-actin in neurological disorders, as presented here, is an important feature of TrJ/+ pathology as a Charcot-Marie-Tooth (CMT) model. Copyright © 2012 Wiley Periodicals, Inc.

  13. Actinic Mask Inspection at the ALS Initial Design Review

    International Nuclear Information System (INIS)

    Barty, A; Chapman, H; Sweeney, D; Levesque, R; Bokor, J; Gullikson, E; Jong, S; Liu, Y; Yi, M; Denbeaux, G; Goldberg, K; Naulleau, P; Denham, P; Rekawa, S; Baston, P; Tackaberry, R; Barale, P

    2003-01-01

    This report is the first milestone report for the actinic mask blank inspection project conducted at the VNL, which forms sub-section 3 of the Q1 2003 mask blank technology transfer program at the VNL. Specifically this report addresses deliverable 3.1.1--design review and preliminary tool design. The goal of this project is to design an actinic mask inspection tool capable of operating in two modes: high-speed scanning for the detection of multilayer defects (inspection mode), and a high-resolution aerial image mode in which the image emulates the imaging illumination conditions of a stepper system (aerial image or AIM mode). The purpose and objective of these two modes is as follows: (1) Defect inspection mode--This imaging mode is designed to scan large areas of the mask for defects EUV multilayer coatings. The goal is to detect the presence of multilayer defects on a mask blank and to store the co-ordinates for subsequent review in AIM mode, thus it is not essential that the illumination and imaging conditions match that of a production stepper. Potential uses for this imaging mode include: (a) Correlating the results obtained using actinic inspection with results obtained using other non-EUV defect inspection systems to verify that the non-EUV scanning systems are detecting all critical defects; (b) Gaining sufficient information to associate defects with particular processes, such as various stages of the multilayer deposition or different modes of operation of the deposition tool; and (c) Assessing the density and EUV impact of surface and multilayer anomalies. Because of the low defect density achieved using current multilayer coating technology it is necessary to be able to efficiently scan large areas of the mask in order to obtain sufficient statistics for use in cross-correlation experiments. Speed of operation as well as sensitivity is therefore key to operation in defect inspection mode. (2) Aerial Image Microscope (AIM) mode--In AIM mode the tool is

  14. Studio della superficie degli impianti dentali in titanio: La nanotecnologia nella valutazione delle nuove superfici implantari in rapporto all'osteointegrazione dei mascellari

    OpenAIRE

    Ricci, Emanuele

    2015-01-01

    Il presente lavoro parte dalla descrizione dei processi di rimodellamento osseo mascellare a seguito della perdita di elementi dentari e la successiva riabilitazione mediante impianto dentale osteointegrato. Approfondiremo proprio i complessi aspetti dell’osteointegrazione su superfici implantari in titanio sia a livello micro che macroscopico. Nel campo dell’implantologia, infatti, il titanio risulta essere il materiale maggiormente impiegato in virtù della sua eccellente biocompatibilità...

  15. Hyperosmotic stress induces Rho/Rho kinase/LIM kinase-mediated cofilin phosphorylation in tubular cells: key role in the osmotically triggered F-actin response

    DEFF Research Database (Denmark)

    Thirone, Ana C P; Speight, Pam; Zulys, Matthew

    2009-01-01

    Hyperosmotic stress induces cytoskeleton reorganization and a net increase in cellular F-actin, but the underlying mechanisms are incompletely understood. While de novo F-actin polymerization likely contributes to the actin response, the role of F-actin severing is unknown. To address this proble...

  16. Superficial x-ray in-vivo dosimetry with MOSFET detectors

    International Nuclear Information System (INIS)

    Cheung, T.; Yu, P.K.N.; Butson, M.J.; Cancer Services, Wollongong, NSW

    2004-01-01

    Full text: This note investigates in-vivo dosimetry using a Metal Oxide Semiconductor Field Effect Transistor (MOSFET) for radiotherapy treatment at superficial and orthovoltage x-ray energies. This was performed within one fraction of the patient's treatment. Standard measurements along with energy response of the detector are given. Results showed that the MOSFET measurements in-vivo agreed with calculated results on average within ± 5.6% over all superficial and orthovoltage energies. These variations were slightly larger than TLD results with variations between measured and calculated results being ± 5.0% for the same patient measurements. The MOSFET device provides adequate in-vivo dosimetry for superficial and orthovoltage energy treatments with the accuracy of the measurements seeming to be relatively on par with TLD in our case. The MOSFET does have the advantage of returning a relatively immediate dosimetric result after irradiation. Copyright (2004) Australasian College of Physical Scientists and Engineers in Medicine

  17. Castelli di carta. La piega per la costruzione di superfici articolate

    Directory of Open Access Journals (Sweden)

    Andrea Casale

    2012-06-01

    Full Text Available L’antica arte del piegare la carta, l’Origami, sta vivendo un rinnovato interesse che coinvolge molti aspetti della ricerca.  Con il termine origami, si intende lo studio del modo di piegare il foglio di carta per imporgli una specifica forma.  La superficie tassellata per mezzo di specifiche pieghe, si propone come un nuovo soggetto di studio: la superficie piegata articolata. La forma congiunta al movimento, assume un particolare interesse nella contemporanea ricerca geometrica e architettonica. Il panorama contemporaneo, propone opere di architettura “responsiva”, capaci di modificare le proprie caratteristiche per adeguarsi a nuove condizioni. La superficie piegata articolata sembra particolarmente adatta a descrivere questo modo d’intendere l’architettura, reagendo a diverse volontà e di conseguenza modificando la propria conformazione attraverso un attento controllo progettuale della forma.

  18. Neurothekeoma palpebrae in association with multiple superficial angiomyxomas: Tegumental Angiomyxoma- Neurothekeoma syndrome (TAN syndrome

    Directory of Open Access Journals (Sweden)

    Tan Aik Kah

    2011-09-01

    Full Text Available We report a case of 10-year-old Indian girl with history of multiple superficial angiomyxoma, presented with three months history of painless right upper lid swelling. There were no visual dysfunctions. Previously, the patient had multiple superficial angiomyxoma (left pinna, left upper cheek, left upper limb, chest, right axilla, hard palate and epidermal cyst (chin. The histopathological specimens were negative to S-100 protein antibody. Systemic review and family history was unremarkable. Excision biopsy and upper lid reconstruction were performed. Intraoperatively the tumor was multilobulated, firm, well encapsulated and did not invade the underlying tarsal plate. Histopathological features of the upperlid tumor were consistent with nerves sheath myxoma (neurothekeoma. To the best of the authors’ knowledge, this is the first reported case of neurothekeoma in association with multiple superficial angiomyxoma.

  19. Neurothekeoma palpebrae in association with multiple superficial angiomyxomas: Tegumental Angiomyxoma-Neurothekeoma syndrome (TAN syndrome).

    Science.gov (United States)

    Kah, Tan Aik; Yong, Ku Chui; Annuar, Faridah Hanom

    2011-07-01

    We report a case of 10-year-old Indian girl with history of multiple superficial angiomyxoma, presented with three months history of painless right upper lid swelling. There were no visual dysfunctions. Previously, the patient had multiple superficial angiomyxoma (left pinna, left upper cheek, left upper limb, chest, right axilla, hard palate) and epidermal cyst (chin). The histopathological specimens were negative to S-100 protein antibody. Systemic review and family history was unremarkable. Excision biopsy and upper lid reconstruction were performed. Intraoperatively the tumor was multilobulated, firm, well encapsulated and did not invade the underlying tarsal plate. Histopathological features of the upperlid tumor were consistent with nerves sheath myxoma (neurothekeoma). To the best of the authors' knowledge, this is the first reported case of neurothekeoma in association with multiple superficial angiomyxoma.

  20. Effects of pressure drop and superficial velocity on the bubbling fluidized bed incinerator.

    Science.gov (United States)

    Wang, Feng-Jehng; Chen, Suming; Lei, Perng-Kwei; Wu, Chung-Hsing

    2007-12-01

    Since performance and operational conditions, such as superficial velocity, pressure drop, particles viodage, and terminal velocity, are difficult to measure on an incinerator, this study used computational fluid dynamics (CFD) to determine numerical solutions. The effects of pressure drop and superficial velocity on a bubbling fluidized bed incinerator (BFBI) were evaluated. Analytical results indicated that simulation models were able to effectively predict the relationship between superficial velocity and pressure drop over bed height in the BFBI. Second, the models in BFBI were simplified to simulate scale-up beds without excessive computation time. Moreover, simulation and experimental results showed that minimum fluidization velocity of the BFBI must be controlled in at 0.188-3.684 m/s and pressure drop was mainly caused by bed particles.

  1. Superficie Específica y Estado Vital de la Materia

    OpenAIRE

    J. Hernando Ordoñez

    1992-01-01

    Se propone reemplazar el término de materia viva por el de estado vital de la materia por considerarlo adecuado, pues la materia es la misma, lo que cambia es su estado. Se estudia la relación entre la superficie específica y el estado vital de la materia. Constante del estado vital y el valor crítico de la superficie específica. Qué es la fuerza de inducción vital? Diferenciación funcional de las células. Cómo regula la unidad biológica su superficie específica? La materia que piens...

  2. "A Tale of Two Planes": Deep Versus Superficial Serratus Plane Block for Postmastectomy Pain Syndrome.

    Science.gov (United States)

    Piracha, Mohammad M; Thorp, Stephen L; Puttanniah, Vinay; Gulati, Amitabh

    Postmastectomy pain syndrome (PMPS) is a significant burden for breast cancer survivors. Although multiple therapies have been described, an evolving field of serratus anterior plane blocks has been described in this population. We describe the addition of the deep serratus anterior plane block (DSPB) for PMPS. Four patients with history of PMPS underwent DSPB for anterior chest wall pain. A retrospective review of these patients' outcomes was obtained through postprocedure interviews. Three of the patients previously had a superficial serratus anterior plane block, which was not as efficacious as the DSPB. The fourth patient had a superficial serratus anterior plane that was difficult to separate with hydrodissection but had improved pain control with a DSPB. We illustrate 4 patients who have benefitted from a DSPB and describe indications that this block may be more efficacious than a superficial serratus plane block. Further study is recommended to understand the intercostal nerve branches within the lateral and anterior muscular chest wall planes.

  3. MOSFET dosimetry in-vivo at superficial and orthovoltage x-ray energies

    International Nuclear Information System (INIS)

    Cheung, T.; Yu, P.K.N.; Butson, M.J.; Illawarra Cancer Care Centre, Wollongong, NSW

    2003-01-01

    This note investigates in-vivo dosimetry using a Metal Oxide Semiconductor Field Effect Transistor (MOSFET) for radiotherapy treatment at superficial and orthovoltage x-ray energies. This was performed within one fraction of the patients treatment. Standard measurements along with energy response of the detector are given. Results showed that the MOSFET measurements in-vivo agreed with calculated results on average within ± 5.6% over all superficial and orthovoltage energies. These variations were slightly larger than TLD results with variations between measured and calculated results being ± 5.0% for the same patient measurements. The MOSFET device provides adequate in-vivo dosimetry for superficial and orthovoltage energy treatments with the accuracy of the measurements seeming to be relatively on par with TLD in our case. The MOSFET does have the advantage of returning a relatively immediate dosimetric result after irradiation. Copyright (2003) Australasian College of Physical Scientists and Engineers in Medicine

  4. Prevalence and causative agents of superficial mycoses in a textile factory in Adana, Turkey.

    Science.gov (United States)

    Celik, E; Ilkit, M; Tanir, F

    2003-09-01

    This study was carried out in a textile factory settled in the city center of Adana, Turkey. The workers were evaluated for the presence of superficial mycoses and the interaction of their working environment or working condition. A total of 431 textile workers were included in the study, with a male to female ratio of 378 (87.7%) to 53 (12.3%) and an age range of 19-52 (mean: 33.7 +/- 6.8). Direct examination and/or culture revealed superficial mycoses in 73 (16.9%) workers, among them 56 (76.7%) were classified as dermatophytoses, 8 (11.0%) as Pityriasis versicolor while in nine (12.3%) of the cases, no causative agent could be determined. Trichophyton rubrum (57.1%) and T. mentagrophytes (42.9%) were the two species isolated on culture. This study emphasized that textile workers should be admitted as a risk group for superficial mycoses, especially tinea pedis.

  5. Investigation of Superficial Resistance of Different Purity Copper at Boiling Nitrogen Temperature Depending on Treatment of Current-Conducting Layer

    International Nuclear Information System (INIS)

    Kutovoj, V.A.; Nikolaenko, A.A.; Stoev, P.I.

    2007-01-01

    Results of this scientific work show influence of annealing temperature and deformation degree of initial MOB copper and after electron beam refining on superficial resistance at temperature of boiling nitrogen. It is shown, that 30 % deformation and annealing in 873...923 K temperature range results in appreciable reduction of superficial resistance at the investigated samples of copper. The lowest values of superficial resistance after thermal and mechanical treatment were observed in the samples after electron beam refinement

  6. Coronin 3 involvement in F-actin-dependent processes at the cell cortex

    International Nuclear Information System (INIS)

    Rosentreter, Andre; Hofmann, Andreas; Xavier, Charles-Peter; Stumpf, Maria; Noegel, Angelika A.; Clemen, Christoph S.

    2007-01-01

    The actin interaction of coronin 3 has been mainly documented by in vitro experiments. Here, we discuss coronin 3 properties in the light of new structural information and focus on assays that reflect in vivo roles of coronin 3 and its impact on F-actin-associated functions. Using GFP-tagged coronin 3 fusion proteins and RNAi silencing we show that coronin 3 has roles in wound healing, protrusion formation, cell proliferation, cytokinesis, endocytosis, axonal growth, and secretion. During formation of cell protrusions actin accumulation precedes the focal enrichment of coronin 3 suggesting a role for coronin 3 in events that follow the initial F-actin assembly. Moreover, we show that coronin 3 similar to other coronins interacts with the Arp2/3-complex and cofilin indicating that this family in general is involved in regulating Arp2/3-mediated events

  7. Intensified photodynamic therapy of actinic keratoses with fractional CO2 laser

    DEFF Research Database (Denmark)

    Togsverd-Bo, K; Haak, C S; Thaysen-Petersen, D

    2012-01-01

    Photodynamic therapy (PDT) with methyl aminolaevulinate (MAL) is effective for thin actinic keratoses (AKs) in field-cancerized skin. Ablative fractional laser resurfacing (AFXL) creates vertical channels that facilitate MAL uptake and may improve PDT efficacy....

  8. The effect of ionizing radiation on the filamentous actin of vascular endothelial cell

    International Nuclear Information System (INIS)

    Yao Xiaowu; Chen Shisheng; Yang Lihe; Lin Juelong; Yang Haiwei

    2006-01-01

    Objective: To observe the ionizing radiation effect on filamentous actin of vascular endothelial cell and explore its mechanism. Methods: The vascular endothelial cells were irradiated with 0, 2, 4, 6, 8, 10 and 12 Gy 60 Co γ-rays. The cytoskeleton was observed with CLSM at 6 hs after the irradiation and the cytoskeleton protein F-actin detected with flow cytometry after 12 and 24 hs. Results: The damage to cytoskeletons increased with the radiation dose. The cytoskeleton protein F-actin was significantly decreased at 12 hs after the irradiation, and then recovered after 24 hs. Conclusion: Ionizing radiation caused vascular endothelial cell injury by damaging the cytoskeleton and depolymerizating the F-actin. (authors)

  9. The interaction between the adaptor protein APS and Enigma is involved in actin organisation

    DEFF Research Database (Denmark)

    Barres, Romain; Gonzalez, Teresa; Le Marchand-Brustel, Yannick

    2005-01-01

    that was previously shown to be associated with the actin cytoskeleton. In HEK 293 cells, Enigma interacted specifically with APS, but not with the APS-related protein SH2-B. This interaction required the NPTY motif of APS and the LIM domains of Enigma. In NIH-3T3 cells that express the insulin receptor, Enigma...... cytoskeleton organisation, expression of Enigma alone led to the formation of F-actin clusters. Similar alteration in actin cytoskeleton organisation was observed in cells expressing both Enigma and APS with a mutation in the NPTY motif. These results identify Enigma as a novel APS-binding protein and suggest...... that the APS/Enigma complex plays a critical role in actin cytoskeleton organisation....

  10. Cofilin phosphorylation is elevated after F-actin disassembly induced by Rac1 depletion

    DEFF Research Database (Denmark)

    Liu, Linna; Li, Jing; Zhang, Liwang

    2015-01-01

    Cytoskeletal reorganization is essential to keratinocyte function. Rac1 regulates cytoskeletal reorganization through signaling pathways such as the cofilin cascade. Cofilin severs actin filaments after activation by dephosphorylation. Rac1 was knocked out in mouse keratinocytes and it was found...... that actin filaments disassembled. In the epidermis of mice in which Rac1 was knocked out only in keratinocytes, cofilin phosphorylation was aberrantly elevated, corresponding to repression of the phosphatase slingshot1 (SSH1). These effects were independent of the signaling pathways for p21-activated kinase....../LIM kinase (Pak/LIMK), protein kinase C, or protein kinase D or generation of reactive oxygen species. Similarly, when actin polymerization was specifically inhibited or Rac1 was knocked down, cofilin phosphorylation was enhanced and SSH1 was repressed. Repression of SSH1 partially blocked actin...

  11. Mena–GRASP65 interaction couples actin polymerization to Golgi ribbon linking

    Science.gov (United States)

    Tang, Danming; Zhang, Xiaoyan; Huang, Shijiao; Yuan, Hebao; Li, Jie; Wang, Yanzhuang

    2016-01-01

    In mammalian cells, the Golgi reassembly stacking protein 65 (GRASP65) has been implicated in both Golgi stacking and ribbon linking by forming trans-oligomers through the N-terminal GRASP domain. Because the GRASP domain is globular and relatively small, but the gaps between stacks are large and heterogeneous, it remains puzzling how GRASP65 physically links Golgi stacks into a ribbon. To explore the possibility that other proteins may help GRASP65 in ribbon linking, we used biochemical methods and identified the actin elongation factor Mena as a novel GRASP65-binding protein. Mena is recruited onto the Golgi membranes through interaction with GRASP65. Depleting Mena or disrupting actin polymerization resulted in Golgi fragmentation. In cells, Mena and actin were required for Golgi ribbon formation after nocodazole washout; in vitro, Mena and microfilaments enhanced GRASP65 oligomerization and Golgi membrane fusion. Thus Mena interacts with GRASP65 to promote local actin polymerization, which facilitates Golgi ribbon linking. PMID:26538023

  12. Corte y sellado reversible de dientes para obtener superficies internas de esmalte incólumes

    Directory of Open Access Journals (Sweden)

    Ana Milena Santiago-Medina

    2015-07-01

    Full Text Available Las superficies internas de los dientes, que se obtienen por cortes con discos, están generalmente desorganizadas por los giros, la velocidad y la rugosidad del disco. El calor altera la superficie y el agua de la refrigeración la composición química porque disuelve los componentes. En estudios de difusión, el perfil se borra por el corte, lo que anula resultados. Objetivo: desarrollar un método de sellado de superficies internas, ya cortadas antes de los ensayos de difusión, sin dejar trazas químicas o partículas. Materiales y metedos: se probó in vitro la habilidad de cuatro materiales de sellado con 40 especimenes, provenientes de 20 coronas de terceros molares, cortadas sagitalmente. Los especímenes se distribuyeron aleatoriamente en cuatro grupos (n = 10: G1: Teflón (Topex®, G2: Tela de caucho (Dental DAM®, G3: película de cloruro de polivinilo (Vinilpel® y G4: Cinta aislante (Tesa®. Se usó una solución de azul de metileno (AM al 2% por 6 días como indicadora de filtración. Por observación visual de cada muestra, se le asignó un valor (sí o no dependiendo de la presencia o ausencia de cualquier punto MB sobre la superficie de sellado. Los resultados fueron analizados siguiendo un modelo estadístico de respuesta binomial. Resultados: Siguiendo los procedimientos descritos, el teflón es el único material que previene la filtración de AM en la superficie interna expuesta por el corte. Conclusión: Se propone un protocolo de sellado reversible de las superficies internas con teflón, previo al corte de los dientes, para no alterar los resultados experimentales.

  13. Evaluation of anatomy and variations of superficial palmar arch and upper extremity arteries with CT angiography.

    Science.gov (United States)

    Kaplanoglu, Hatice; Beton, Osman

    2017-04-01

    To evaluate the abnormalities and variations of the arterial system of upper extremities and superficial palmar arch with computed tomography angiography and to guide the clinician during this procedure. A total of 156 upper extremities of 78 cases were retrospectively analyzed using computed tomography angiography. The study was approved by the local ethics committee of the hospital. From the analysis of the computed tomography angiography images, the following information was recorded; the diameters and abnormalities of radial, ulnar and brachial arteries in both upper extremities, the presence of atherosclerotic changes or stenosis in these arteries, whether the superficial palmar arch was complete or incomplete, and arterial dominance. Also, the computed tomography angiography classification of superficial palmar arch distribution and anatomic configuration was performed. The mean baseline diameters of the radial, ulnar and brachial arteries of the cases were; 2.8 ± 0.6, 2.5 ± 0.7, and 4.7 ± 0.6 mm, respectively. A complete superficial palmar arch was observed in 69.2 % of the right hands and 70.5 % of the left hands. For the superficial palmar arches on the right side, the radial artery was dominant in two and the ulnar artery was dominant in 47 with the remaining showing codominance. On the left side, the radial artery was dominant in one hand, with the ulnar artery being dominant in 49 cases, and in 28 cases, there was codominance. In the superficial palmar arch classification, four of the arches (A-D) were defined as complete and the remaining three (E-G) as incomplete. The current study clarified different variations in palmar circulation and forearm arteries to aid the surgeon during trans-radial or trans-ulnar catheterization, hemodialysis, or coronary artery bypass grafting.

  14. Ultrasound of musculoskeletal soft-tissue tumors superficial to the investing fascia.

    Science.gov (United States)

    Hung, Esther Hiu Yee; Griffith, James Francis; Ng, Alex Wing Hung; Lee, Ryan Ka Lok; Lau, Domily Ting Yi; Leung, Jason Chi Shun

    2014-06-01

    The objective of our study was to evaluate the diagnostic accuracy of ultrasound in assessing musculoskeletal soft-tissue tumors superficial to the investing fascia. Seven hundred fourteen superficial soft-tissue tumors evaluated with ultrasound by two musculoskeletal radiologists were retrospectively reviewed. In all ultrasound reports, the reporting radiologists provided one, two, or three diagnoses depending on their perceived level of diagnostic certainty. Two hundred forty-seven tumors had subsequent histologic correlation, thus allowing the accuracy of the ultrasound diagnosis to be determined. Images of the lesions with a discordant ultrasound diagnosis and histologic diagnosis were reviewed, and the ultrasound features were further classified as concordant with the known histologic diagnosis, concordant with the known histologic diagnosis with atypical features present, or discordant with the known histologic diagnosis. Four hundred sixty-seven tumors without pathologic confirmation were followed up clinically. Overall the accuracy of ultrasound examination for assessing superficial soft-tissue masses was 79.0% when all differential diagnoses were considered and 77.0% when only the first differential diagnosis was considered. The sensitivity and specificity of the first ultrasound diagnosis were 95.2% and 94.3%, respectively, for lipoma; 73.0% and 97.7% for vascular malformation; 80.0% and 95.4% for epidermoid cyst; and 68.8% and 95.2% for nerve sheath tumor. Reduced observer awareness of specific tumor entities tended to contribute to underdiagnosis more than poor specificity of ultrasound findings. Most tumors (236/247, 96%) were benign. The sensitivity and specificity of ultrasound for identifying malignant superficial soft-tissue tumors was 94.1% and 99.7%, respectively. The diagnostic accuracy of ultrasound in the assessment of superficial musculoskeletal soft-tissue tumors is high and can be improved through increased radiologist awareness of less

  15. Live cell imaging of actin dynamics in dexamethasone-treated porcine trabecular meshwork cells.

    Science.gov (United States)

    Fujimoto, Tomokazu; Inoue, Toshihiro; Inoue-Mochita, Miyuki; Tanihara, Hidenobu

    2016-04-01

    The regulation of the actin cytoskeleton in trabecular meshwork (TM) cells is important for controlling outflow of the aqueous humor. In some reports, dexamethasone (DEX) increased the aqueous humor outflow resistance and induced unusual actin structures, such as cross-linked actin networks (CLAN), in TM cells. However, the functions and dynamics of CLAN in TM cells are not completely known, partly because actin stress fibers have been observed only in fixed cells. We conducted live-cell imaging of the actin dynamics in TM cells with or without DEX treatment. An actin-green fluorescent protein (GFP) fusion construct with a modified insect virus was transfected into porcine TM cells. Time-lapse imaging of live TM cells treated with 25 μM Y-27632 and 100 nM DEX was performed using an inverted fluorescence microscope. Fluorescent images were recorded every 15 s for 30 min after Y-27632 treatment or every 30 min for 72 h after DEX treatment. The GFP-actin was expressed in 22.7 ± 10.9% of the transfected TM cells. In live TM cells, many actin stress fibers were observed before the Y-27632 treatment. Y-27632 changed the cell shape and decreased stress fibers in a time-dependent manner. In fixed cells, CLAN-like structures were seen in 26.5 ± 1.7% of the actin-GFP expressed PTM cells treated with DEX for 72 h. In live imaging, there was 28% CLAN-like structure formation at 72 h after DEX treatment, and the lifetime of CLAN-like structures increased after DEX treatment. The DEX-treated cells with CLAN-like structures showed less migration than DEX-treated cells without CLAN-like structures. Furthermore, the control cells (without DEX treatment) with CLAN-like structures also showed less migration than the control cells without CLAN-like structures. These results suggested that CLAN-like structure formation was correlated with cell migration in TM cells. Live cell imaging of the actin cytoskeleton provides valuable information on the actin dynamics in TM

  16. Concerning the dynamic instability of actin homolog ParM

    International Nuclear Information System (INIS)

    Popp, David; Yamamoto, Akihiro; Iwasa, Mitsusada; Narita, Akihiro; Maeda, Kayo; Maeda, Yuichiro

    2007-01-01

    Using in vitro TIRF- and electron-microscopy, we reinvestigated the dynamics of native ParM, a prokaryotic DNA segregation protein and actin homolog. In contrast to a previous study, which used a cysteine ParM mutant, we find that the polymerization process of wild type ATP-ParM filaments consists of a polymerization phase and a subsequent steady state phase, which is dynamically unstable, like that of microtubules. We find that the apparent bidirectional polymerization of ParM, is not due to the intrinsic nature of this filament, but results from ParM forming randomly oriented bundles in the presence of crowding agents. Our results imply, that in the bacterium, ParM filaments spontaneously form bipolar bundles. Due to their intrinsic dynamic instability, ParM bundles can efficiently 'search' the cytoplasmic lumen for DNA, bind it equally well at the bipolar ends and segregate it approximately symmetrically, by the insertion of ParM subunits at either end

  17. Assembly properties of the Bacillus subtilis actin, MreB.

    Science.gov (United States)

    Mayer, Joshua A; Amann, Kurt J

    2009-02-01

    The bacterial actin MreB has been implicated in a variety of cellular roles including cell shape determination, cell wall synthesis, chromosome condensation and segregation, and the establishment and maintenance of cell polarity. Toward elucidating a clearer understanding of how MreB functions inside the bacterial cell, we investigated biochemically the polymerization of MreB from Bacillus subtilis. Light scattering and sedimentation assays revealed pH-, ionic-, cationic-, and temperature-dependent behavior. B. subtilis MreB polymerizes in the presence of millimolar divalent cations in a protein concentration-dependent manner. Polymerization is favored by decreasing pH and inhibited by monovalent salts and low temperatures. Although B. subtilis MreB binds and hydrolyzes both ATP and GTP, it does not require a bound nucleotide for assembly and polymerizes indistinguishably regardless of the nucleotide species bound, with a critical concentration of approximately 900 nM. A number of the presently reported properties of B. subtilis MreB differ significantly from those of T. maritima MreB1 (Bean and Amann [2008]: Biochemistry 47: 826-835), including the nucleotide requirements and temperature and ionic effects on polymerization state. These observations collectively suggest that additional factors interact with MreB to account for its complex dynamic behavior in cells.

  18. Actinic Keratosis Clinical Practice Guidelines: An Appraisal of Quality

    Directory of Open Access Journals (Sweden)

    Joslyn S. Kirby

    2015-01-01

    Full Text Available Actinic keratosis (AK is a common precancerous skin lesion and many AK management guidelines exist, but there has been limited investigation into the quality of these documents. The objective of this study was to assess the strengths and weaknesses of guidelines that address AK management. A systematic search for guidelines with recommendations for AK was performed. The Appraisal of Guidelines for Research and Evaluation (AGREE II was used to appraise the quality of guidelines. Multiple raters independently reviewed each of the guidelines and applied the AGREE II tool and scores were calculated. Overall, 2,307 citations were identified and 7 fulfilled the study criteria. The Cancer Council of Australia/Australian Cancer Network guideline had the highest mean scores and was the only guideline to include a systematic review, include an evidence rating for recommendations, and report conflicts of interest and funding sources. High-quality, effective guidelines are evidence-based with recommendations that are concise and organized, so practical application is facilitated. Features such as concise tables, pictorial diagrams, and explicit links to evidence are helpful. However, the rigor and validity of some guidelines were weak. So, it is important for providers to be aware of the features that contribute to a high-quality, practical document.

  19. New and current preventive treatment options in actinic keratosis.

    Science.gov (United States)

    Arenberger, P; Arenbergerova, M

    2017-09-01

    Actinic keratosis (AK) is a characteristic skin lesion on skin areas of subjects with mainly phototype I and phototype II, or with specific genetic factors and who are exposed to prolonged ultraviolet radiation. AK may be considered a precursor of in situ squamous cell carcinoma (SCC), a type of non-melanoma skin cancer (NMSC). However, it is still not possible to predict which AK lesions will develop into SCC. Early treatment of AK is therefore recommended. Despite the increasing number of patients with AK developing into SCC, to date, there is still no clear suggestion of therapeutic strategy for AK. Current treatment consists of a multitude of topical lesion-directed or field-directed therapies or a combination of both. Recently, orally administered nicotinamide has shown to significantly reduce rates of new NMSC and AK in high-risk patients. This study aims to provide an update on the most relevant information about AK and to provide an insight into current and new treatment options. © 2017 European Academy of Dermatology and Venereology.

  20. Force-velocity measurements of a few growing actin filaments.

    Directory of Open Access Journals (Sweden)

    Coraline Brangbour

    2011-04-01

    Full Text Available The polymerization of actin in filaments generates forces that play a pivotal role in many cellular processes. We introduce a novel technique to determine the force-velocity relation when a few independent anchored filaments grow between magnetic colloidal particles. When a magnetic field is applied, the colloidal particles assemble into chains under controlled loading or spacing. As the filaments elongate, the beads separate, allowing the force-velocity curve to be precisely measured. In the widely accepted Brownian ratchet model, the transduced force is associated with the slowing down of the on-rate polymerization. Unexpectedly, in our experiments, filaments are shown to grow at the same rate as when they are free in solution. However, as they elongate, filaments are more confined in the interspace between beads. Higher repulsive forces result from this higher confinement, which is associated with a lower entropy. In this mechanism, the production of force is not controlled by the polymerization rate, but is a consequence of the restriction of filaments' orientational fluctuations at their attachment point.

  1. Bacterias en superficies contactadas durante las tomas radiográficas intraorales

    OpenAIRE

    Lee, Guihan; Calderón-Ubaquí, Victor; Sacsaquispe-Contreras, Sonia

    2016-01-01

    Objetivo: Determinar la presencia de bacterias mediante el análisis microbiológico en las superficies contactadas por el operador durante la toma y procesado de radiografías intraorales en diferentes momentos del día en el Servicio de Radiología Oral de la UPCH. Materiales y métodos: Se realizó un muestreo en nueve superficies del servicio de radiología oral. Las muestras se tomaron en dos momentos por el mismo investigador; al inicio y al finalizar las actividades en el servicio, se realizó ...

  2. Recent advances in high-throughput molecular marker identification for superficial and invasive bladder cancers

    DEFF Research Database (Denmark)

    Andersen, Lars Dyrskjøt; Zieger, Karsten; Ørntoft, Torben Falck

    2007-01-01

    individually contributed to the management of the disease. However, the development of high-throughput techniques for simultaneous assessment of a large number of markers has allowed classification of tumors into clinically relevant molecular subgroups beyond those possible by pathological classification. Here......Bladder cancer is the fifth most common neoplasm in industrialized countries. Due to frequent recurrences of the superficial form of this disease, bladder cancer ranks as one of the most common cancers. Despite the description of a large number of tumor markers for bladder cancers, none have......, we review the recent advances in high-throughput molecular marker identification for superficial and invasive bladder cancers....

  3. Managment of superficial infantile capillary hemangiomas with topical timolol maleate solution.

    Science.gov (United States)

    Rizvi, Syed Ali Raza; Yusuf, Faraz; Sharma, Rajeev; Rizvi, Syed Wajahat Ali

    2015-01-01

    Capillary hemangioma is the most common benign tumor of eyelids and orbit in children. Recently, a topical beta blocker has been reported as an effective treatment for superficial capillary hemangiomas. We present a case report of two children having large capillary hemangiomas who responded well to topical treatment by 0.5% timolol maleate solution. After 12 months of treatment, the lesion has significantly reduced in size, thickness, and color in both cases. Thus, we conclude that long-term use of topical 0.5% timolol maleate solution is safe and effective in treating superficial capillary hemangiomas.

  4. Superficial Temporal Artery Pseudoaneurysm: A Conservative Approach in a Critically Ill Patient

    International Nuclear Information System (INIS)

    Grasso, Rosario Francesco; Quattrocchi, Carlo Cosimo; Crucitti, Pierfilippo; Carboni, Giampiero; Coppola, Roberto; Zobel, Bruno Beomonte

    2007-01-01

    A 71-year-old man affected by cardio- and cerebrovascular disease experienced an accidental fall and trauma to the fronto-temporal area of the head. A few weeks later a growing mass appeared on his scalp. A diagnosis of superficial temporal artery pseudoaneurysm was made following CT and color Doppler ultrasound. His clinical condition favoured a conservative approach by ultrasound-guided compression and subsequent surgical resection. A conservative approach should be considered the treatment of choice in critically ill patients affected by superficial temporal artery pseudoaneurysm

  5. Secondary superficial siderosis of the central nervous system in a patient presenting with sensorineural hearing loss

    International Nuclear Information System (INIS)

    Lemmerling, M.; De Praeter, G.; Mollet, P.; Mortele, K.; Kunnen, M.; Mastenbroek, G.

    1998-01-01

    We present a 50-year-old man who was investigated for sensorineural hearing loss. On MRI of the brain superficial siderosis of the central nervous system was seen, while MRI of the spine revealed an ependymoma of the cauda equina. This case illustrates the importance of performing T2-weighted imaging of the brain and posterior fossa when sensorineural hearing loss is present. Spine imaging is mandatory when superficial siderosis of the brain is diagnosed without identification of a bleeding source in the brain. (orig.)

  6. Harvest Date Influences Superficial Scald Development in Granny Smith Apples During Long Term Storage

    OpenAIRE

    ERKAN, Mustafa; PEKMEZCİ, Mustafa

    2014-01-01

    The effects of harvest dates on superficial scald development and postharvest quality in 'Granny Smith' apples (Malus domestica Borkh.) were investigated. Apples were harvested at 15-day intervals during 2 consecutive years (2000-2001) and stored at 0 °C with 90% relative humidity for 8 months. At the end of the 8 - month storage period plus an additional 1 week at 20 °C the percentage of superficial scald was lower (24.4%) in late harvested (November 15) apples than in ap...

  7. Caracterizacion dosimetrica del equipo de Terapia Superficial Therapax del Hospital Mexico

    International Nuclear Information System (INIS)

    Bernui De Vivanco, Maria Giselle; Rojas Rivas, Jorge; Cardenas Retuerto, Augusto

    2007-01-01

    The Servicio de Radioterapia of the Hospital Mexico in San Jose, Costa Rica has a Superficial Therapy equipment Therapax model DXT 300. It describes the measurements for the dosimetric characterization of the configurations 100 kVp, 25 mA and filter of 2.4 mm of Al, and 250 kVp, 10 mA and filter of 2.5 mm of Cu, taking into account that the uncertainties in the measurements of the superficial therapy equipment are higher than in the rank of the megavoltage [es

  8. Radiación acústica por superficies planas: Aplicación a altavoces

    OpenAIRE

    Alba, Jesús; Ramis, Jaime; Espinosa, Víctor; Sánchez, Víctor

    2003-01-01

    En los sistemas de reproducción sonora es habitual la utilización de altavoces dinámicos. Sin embargo, en aplicaciones específicas, puede ser necesaria y/o conveniente la utilización de altavoces alternativos con superficies planas como diafragma, como los electrostáticos o los basados en la tecnología NXT© . Estos altavoces generan el campo acústico mediante la vibración de una superficie rectangular. Se puede suponer, en una primera aproximación, que todos los puntos sobre su su...

  9. Actin dynamics at focal adhesions: a common endpoint and putative therapeutic target for proteinuric kidney diseases.

    Science.gov (United States)

    Sever, Sanja; Schiffer, Mario

    2018-06-01

    Proteinuria encompasses diverse causes including both genetic diseases and acquired forms such as diabetic and hypertensive nephropathy. The basis of proteinuria is a disturbance in size selectivity of the glomerular filtration barrier, which largely depends on the podocyte: a terminally differentiated epithelial cell type covering the outer surface of the glomerulus. Compromised podocyte structure is one of the earliest signs of glomerular injury. The phenotype of diverse animal models and podocyte cell culture firmly established the essential role of the actin cytoskeleton in maintaining functional podocyte structure. Podocyte foot processes, actin-based membrane extensions, contain 2 molecularly distinct "hubs" that control actin dynamics: a slit diaphragm and focal adhesions. Although loss of foot processes encompasses disassembly of slit diaphragm multiprotein complexes, as long as cells are attached to the glomerular basement membrane, focal adhesions will be the sites in which stress due to filtration flow is counteracted by forces generated by the actin network in foot processes. Numerous studies within last 20 years have identified actin binding and regulatory proteins as well as integrins as essential components of signaling and actin dynamics at focal adhesions in podocytes, suggesting that some of them may become novel, druggable targets for proteinuric kidney diseases. Here we review evidence supporting the idea that current treatments for chronic kidney diseases beneficially and directly target the podocyte actin cytoskeleton associated with focal adhesions and suggest that therapeutic reagents that target the focal adhesion-regulated actin cytoskeleton in foot processes have potential to modernize treatments for chronic kidney diseases. Copyright © 2018 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.

  10. Sensory role of actin in auxin-dependent responses of tobacco BY-2.

    Science.gov (United States)

    Huang, Xiang; Maisch, Jan; Nick, Peter

    2017-11-01

    Polar auxin transport depends on the polar localization of auxin-efflux carriers. The cycling of these carriers between cell interior and plasma membrane depends on actin. The dynamic of actin not only affects auxin transport, but also changes the auxin-responsiveness. To study the potential link between auxin responsiveness and actin dynamics, we investigated developmental responses of the non-transformed BY-2 (Nicotiana tabacum L. cv Bright Yellow 2) cell line and the transgenic BY-2 strain GF11 (stably transformed BY-2 cells with a GFP-fimbrin actin-binding domain 2 construct). The developmental process was divided into three distinct stages: cell cycling, cell elongation and file disintegration. Several phenotypes were measured to monitor the cellular responses to different concentrations of exogenous natural auxin (Indole-3-acetic acid, IAA). We found that auxin stimulated and prolonged the mitotic activity, and delayed the exit from the proliferation phase. However, both responses were suppressed in the GF11 line. At the stationary phase of the cultivation cycle, auxin strongly accelerated the cell file disintegration. Interestingly, it was not suppressed but progressed to a more complete disintegration in the GF11 line. During the cultivation cycle, we also followed the organization of actin in the GF11 line and did not detect any significant difference in actin organization from untreated control or exogenous IAA treatment. Therefore, our findings indicate that the specific differences observed in the GF11 line must be linked with a function of actin that is not structural. It means that there is a sensory role of actin for auxin signaling. Copyright © 2017 Elsevier GmbH. All rights reserved.

  11. The actin homologue MreB organizes the bacterial cell membrane

    OpenAIRE

    Strahl, Henrik; Bürmann, Frank; Hamoen, Leendert W.

    2014-01-01

    The eukaryotic cortical actin cytoskeleton creates specific lipid domains, including lipid rafts, which determine the distribution of many membrane proteins. Here we show that the bacterial actin homologue MreB displays a comparable activity. MreB forms membrane-associated filaments that coordinate bacterial cell wall synthesis. We noticed that the MreB cytoskeleton influences fluorescent staining of the cytoplasmic membrane. Detailed analyses combining an array of mutants, using specific lip...

  12. Spatiotemporal dynamics of actin remodeling and endomembrane trafficking in alveolar epithelial type I cell wound healing.

    Science.gov (United States)

    Godin, Lindsay M; Vergen, Jorge; Prakash, Y S; Pagano, Richard E; Hubmayr, Rolf D

    2011-04-01

    Alveolar epithelial type I cell (ATI) wounding is prevalent in ventilator-injured lungs and likely contributes to pathogenesis of "barotrauma" and "biotrauma." In experimental models most wounded alveolar cells repair plasma membrane (PM) defects and survive insults. Considering the force balance between edge energy at the PM wound margins and adhesive interactions of the lipid bilayer with the underlying cytoskeleton (CSK), we tested the hypothesis that subcortical actin depolymerization is a key facilitator of PM repair. Using real-time fluorescence imaging of primary rat ATI transfected with a live cell actin-green fluorescent protein construct (Lifeact-GFP) and loaded with N-rhodamine phosphatidylethanolamine (PE), we examined the spatial and temporal coordination between cytoskeletal remodeling and PM repair following micropuncture. Membrane integrity was inferred from the fluorescence intensity profiles of the cytosolic label calcein AM. Wounding led to rapid depolymerization of the actin CSK near the wound site, concurrent with accumulation of endomembrane-derived N-rhodamine PE. Both responses were sustained until PM integrity was reestablished, which typically occurs between ∼10 and 40 s after micropuncture. Only thereafter did the actin CSK near the wound begin to repolymerize, while the rate of endomembrane lipid accumulation decreased. Between 60 and 90 s after successful PM repair, after translocation of the actin nucleation factor cortactin, a dense actin fiber network formed. In cells that did not survive micropuncture injury, actin remodeling did not occur. These novel results highlight the importance of actin remodeling in ATI cell repair and suggest molecular targets for modulating the repair process.

  13. The RhoGAP Stard13 controls insulin secretion through F-actin remodeling

    Directory of Open Access Journals (Sweden)

    Heike Naumann

    2018-02-01

    Full Text Available Objective: Actin cytoskeleton remodeling is necessary for glucose-stimulated insulin secretion in pancreatic β-cells. A mechanistic understanding of actin dynamics in the islet is paramount to a better comprehension of β-cell dysfunction in diabetes. Here, we investigate the Rho GTPase regulator Stard13 and its role in F-actin cytoskeleton organization and islet function in adult mice. Methods: We used Lifeact-EGFP transgenic animals to visualize actin cytoskeleton organization and dynamics in vivo in the mouse islets. Furthermore, we applied this model to study actin cytoskeleton and insulin secretion in mutant mice deleted for Stard13 selectively in pancreatic cells. We isolated transgenic islets for 3D-imaging and perifusion studies to measure insulin secretion dynamics. In parallel, we performed histological and morphometric analyses of the pancreas and used in vivo approaches to study glucose metabolism in the mouse. Results: In this study, we provide the first genetic evidence that Stard13 regulates insulin secretion in response to glucose. Postnatally, Stard13 expression became restricted to the mouse pancreatic islets. We showed that Stard13 deletion results in a marked increase in actin polymerization in islet cells, which is accompanied by severe reduction of insulin secretion in perifusion experiments. Consistently, Stard13-deleted mice displayed impaired glucose tolerance and reduced glucose-stimulated insulin secretion. Conclusions: Taken together, our results suggest a previously unappreciated role for the RhoGAP protein Stard13 in the interplay between actin cytoskeletal remodeling and insulin secretion. Keywords: F-actin, Insulin secretion, Islet, Pancreas, Lifeact, Stard13

  14. Electron tomography of cryofixed, isometrically contracting insect flight muscle reveals novel actin-myosin interactions.

    Directory of Open Access Journals (Sweden)

    Shenping Wu

    2010-09-01

    Full Text Available Isometric muscle contraction, where force is generated without muscle shortening, is a molecular traffic jam in which the number of actin-attached motors is maximized and all states of motor action are trapped with consequently high heterogeneity. This heterogeneity is a major limitation to deciphering myosin conformational changes in situ.We used multivariate data analysis to group repeat segments in electron tomograms of isometrically contracting insect flight muscle, mechanically monitored, rapidly frozen, freeze substituted, and thin sectioned. Improved resolution reveals the helical arrangement of F-actin subunits in the thin filament enabling an atomic model to be built into the thin filament density independent of the myosin. Actin-myosin attachments can now be assigned as weak or strong by their motor domain orientation relative to actin. Myosin attachments were quantified everywhere along the thin filament including troponin. Strong binding myosin attachments are found on only four F-actin subunits, the "target zone", situated exactly midway between successive troponin complexes. They show an axial lever arm range of 77°/12.9 nm. The lever arm azimuthal range of strong binding attachments has a highly skewed, 127° range compared with X-ray crystallographic structures. Two types of weak actin attachments are described. One type, found exclusively in the target zone, appears to represent pre-working-stroke intermediates. The other, which contacts tropomyosin rather than actin, is positioned M-ward of the target zone, i.e. the position toward which thin filaments slide during shortening.We present a model for the weak to strong transition in the myosin ATPase cycle that incorporates azimuthal movements of the motor domain on actin. Stress/strain in the S2 domain may explain azimuthal lever arm changes in the strong binding attachments. The results support previous conclusions that the weak attachments preceding force generation are very

  15. Electron Tomography of Cryofixed, Isometrically Contracting Insect Flight Muscle Reveals Novel Actin-Myosin Interactions

    International Nuclear Information System (INIS)

    Wu, Shenping; Liu, Jun; Reedy, Mary C.; Tregear, Richard T.; Winkler, Hanspeter; Franzini-Armstrong, Clara; Sasaki, Hiroyuki; Lucaveche, Carmen; Goldman, Yale E.; Reedy, Michael K.; Taylor, Kenneth A.

    2010-01-01

    Isometric muscle contraction, where force is generated without muscle shortening, is a molecular traffic jam in which the number of actin-attached motors is maximized and all states of motor action are trapped with consequently high heterogeneity. This heterogeneity is a major limitation to deciphering myosin conformational changes in situ. We used multivariate data analysis to group repeat segments in electron tomograms of isometrically contracting insect flight muscle, mechanically monitored, rapidly frozen, freeze substituted, and thin sectioned. Improved resolution reveals the helical arrangement of F-actin subunits in the thin filament enabling an atomic model to be built into the thin filament density independent of the myosin. Actin-myosin attachments can now be assigned as weak or strong by their motor domain orientation relative to actin. Myosin attachments were quantified everywhere along the thin filament including troponin. Strong binding myosin attachments are found on only four F-actin subunits, the 'target zone', situated exactly midway between successive troponin complexes. They show an axial lever arm range of 77 o /12.9 nm. The lever arm azimuthal range of strong binding attachments has a highly skewed, 127 o range compared with X-ray crystallographic structures. Two types of weak actin attachments are described. One type, found exclusively in the target zone, appears to represent pre-working-stroke intermediates. The other, which contacts tropomyosin rather than actin, is positioned M-ward of the target zone, i.e. the position toward which thin filaments slide during shortening. We present a model for the weak to strong transition in the myosin ATPase cycle that incorporates azimuthal movements of the motor domain on actin. Stress/strain in the S2 domain may explain azimuthal lever arm changes in the strong binding attachments. The results support previous conclusions that the weak attachments preceding force generation are very different from

  16. Realignment process of actin stress fibers in single living cells studied by focused femtosecond laser irradiation

    OpenAIRE

    Yasukuni, Ryohei; Spitz, Jean-Alexis; Meallet-Renault, Rachel; Negishi, Takayuki; Tada, Takuji; Hosokawa, Yoichiroh; Asahi, Tsuyoshi; Shukunami, Chisa; Hiraki, Yuji; Masuhara, Hiroshi

    2007-01-01

    Three-dimensional dissection of a single actin stress fiber in a living cell was performed based on multi-photon absorption of a focused femtosecond laser pulse. The realignment process of an actin stress fiber was investigated after its direct cutting by a single-shot femtosecond laser pulse irradiation by high-speed transmission and fluorescence imaging methods. It was confirmed that mechanical force led by the femtosecond laser cutting propagates to entire cell through the cytockelton in a...

  17. Espins are multifunctional actin cytoskeletal regulatory proteins in the microvilli of chemosensory and mechanosensory cells

    Science.gov (United States)

    Sekerková, Gabriella; Zheng, Lili; Loomis, Patricia A.; Changyaleket, Benjarat; Whitlon, Donna S.; Mugnaini, Enrico; Bartles, James R.

    2010-01-01

    Espins are associated with the parallel actin bundles of hair cell stereocilia and are the target of mutations that cause deafness and vestibular dysfunction in mice and humans. Here, we report that espins are also concentrated in the microvilli of a number of other sensory cells: vomeronasal organ sensory neurons, solitary chemoreceptor cells, taste cells and Merkel cells. Moreover, we show that hair cells and these other sensory cells contain novel espin isoforms that arise from a different transcriptional start site and differ significantly from other espin isoforms in their complement of ligand-binding activities and their effects on actin polymerization. The novel espin isoforms of sensory cells bundled actin filaments with high affinity in a Ca2+-resistant fashion, bound actin monomer via a WASP homology 2 domain, bound profilin via a single proline-rich peptide, and caused a dramatic elongation of microvillus-type parallel actin bundles in transfected epithelial cells. In addition, the novel espin isoforms of sensory cells differed from other espin isoforms in that they potently inhibited actin polymerization in vitro, did not bind the Src homology 3 domain of the adapter protein insulin receptor substrate p53 and did not bind the acidic, signaling phospholipid phosphatidylinositol 4,5- bisphosphate. Thus, the espins constitute a family of multifunctional actin cytoskeletal regulatory proteins with the potential to differentially influence the organization, dimensions, dynamics and signaling capabilities of the actin filament-rich, microvillus-type specializations that mediate sensory transduction in a variety of mechanosensory and chemosensory cells. PMID:15190118

  18. Increased actin polymerization and stabilization interferes with neuronal function and survival in the AMPKγ mutant Loechrig.

    Directory of Open Access Journals (Sweden)

    Mandy Cook

    Full Text Available loechrig (loe mutant flies are characterized by progressive neuronal degeneration, behavioral deficits, and early death. The mutation is due to a P-element insertion in the gene for the γ-subunit of the trimeric AMP-activated protein kinase (AMPK complex, whereby the insertion affects only one of several alternative transcripts encoding a unique neuronal isoform. AMPK is a cellular energy sensor that regulates a plethora of signaling pathways, including cholesterol and isoprenoid synthesis via its downstream target hydroxy-methylglutaryl (HMG-CoA reductase. We recently showed that loe interferes with isoprenoid synthesis and increases the prenylation and thereby activation of RhoA. During development, RhoA plays an important role in neuronal outgrowth by activating a signaling cascade that regulates actin dynamics. Here we show that the effect of loe/AMPKγ on RhoA prenylation leads to a hyperactivation of this signaling pathway, causing increased phosphorylation of the actin depolymerizating factor cofilin and accumulation of filamentous actin. Furthermore, our results show that the resulting cytoskeletal changes in loe interfere with neuronal growth and disrupt axonal integrity. Surprisingly, these phenotypes were enhanced by expressing the Slingshot (SSH phosphatase, which during development promotes actin depolymerization by dephosphorylating cofilin. However, our studies suggest that in the adult SSH promotes actin polymerization, supporting in vitro studies using human SSH1 that suggested that SSH can also stabilize and bundle filamentous actin. Together with the observed increase in SSH levels in the loe mutant, our experiments suggest that in mature neurons SSH may function as a stabilization factor for filamentous actin instead of promoting actin depolymerization.

  19. Is chloroplast import of photosynthesis proteins facilitated by an actin-TOC-TIC-VIPP1 complex?

    Science.gov (United States)

    Jouhet, Juliette; Gray, John C

    2009-10-01

    Actin filaments are major components of the cytoskeleton that interact with chloroplast envelope membranes to allow chloroplast positioning and movement, stromule mobility and gravitropism perception. We recently reported that Toc159, a component of the TOC complex of the chloroplast protein import apparatus, interacts directly with actin. The interaction of Toc159 and actin was identified by co-immunoprecipitation and co-sedimentation experiments with detergent-solubilised pea chloroplast envelope membranes. In addition, many of the components of the TOC-TIC protein import apparatus and VIPP1 (vesicle-inducing protein in plastids 1) were identified by mass spectroscopy in the material co-immunoprecipitated with antibodies to actin. Toc159 is the receptor for the import of photosynthesis proteins and VIPP1 is involved in thylakoid membrane formation by inducing vesicle formation from the chloroplast inner envelope membrane, suggesting we may have identified an actin-TOC-TIC-VIPP1 complex that may provide a means of channeling cytosolic preproteins to the thylakoid membrane. The interaction of Toc159 with actin may facilitate exchange between the putative soluble and membrane forms of Toc159 and promote the interaction of cytosolic preproteins with the TOC complex.

  20. Cofilin phosphorylation is elevated after F-actin disassembly induced by Rac1 depletion.

    Science.gov (United States)

    Liu, Linna; Li, Jing; Zhang, Liwang; Zhang, Feng; Zhang, Rong; Chen, Xiang; Brakebusch, Cord; Wang, Zhipeng; Liu, Xinyou

    2015-01-01

    Cytoskeletal reorganization is essential to keratinocyte function. Rac1 regulates cytoskeletal reorganization through signaling pathways such as the cofilin cascade. Cofilin severs actin filaments after activation by dephosphorylation. Rac1 was knocked out in mouse keratinocytes and it was found that actin filaments disassembled. In the epidermis of mice in which Rac1 was knocked out only in keratinocytes, cofilin phosphorylation was aberrantly elevated, corresponding to repression of the phosphatase slingshot1 (SSH1). These effects were independent of the signaling pathways for p21-activated kinase/LIM kinase (Pak/LIMK), protein kinase C, or protein kinase D or generation of reactive oxygen species. Similarly, when actin polymerization was specifically inhibited or Rac1 was knocked down, cofilin phosphorylation was enhanced and SSH1 was repressed. Repression of SSH1 partially blocked actin depolymerization induced by Rac1 depletion. Therefore, aberrant cofilin phosphorylation that induces actin polymerization might be a consequence of actin disassembly induced by the absence of Rac1. © 2015 International Union of Biochemistry and Molecular Biology.

  1. Filament formation of the Escherichia coli actin-related protein, MreB, in fission yeast.

    Science.gov (United States)

    Srinivasan, Ramanujam; Mishra, Mithilesh; Murata-Hori, Maki; Balasubramanian, Mohan K

    2007-02-06

    Proteins structurally related to eukaryotic actins have recently been identified in several prokaryotic organisms. These actin-like proteins (MreB and ParM) and the deviant Walker A ATPase (SopA) play a key role in DNA segregation and assemble into polymers in vitro and in vivo. MreB also plays a role in cellular morphogenesis. Whereas the dynamic properties of eukaryotic actins have been extensively characterized, those of bacterial actins are only beginning to emerge. We have established the fission yeast Schizosaccharomyces pombe as a cellular model for the functional analysis of the Escherichia coli actin-related protein MreB. We show that MreB organizes into linear bundles that grow in a symmetrically bidirectional manner at 0.46 +/- 0.03 microm/min, with new monomers and/or oligomers being added along the entire length of the bundle. Organization of linear arrays was dependent on the ATPase activity of MreB, and their alignment along the cellular long axis was achieved by sliding along the cortex of the cylindrical part of the cell. The cell ends appeared to provide a physical barrier for bundle elongation. These experiments provide new insights into the mechanism of assembly and organization of the bacterial actin cytoskeleton.

  2. The assembly of MreB, a prokaryotic homolog of actin.

    Science.gov (United States)

    Esue, Osigwe; Cordero, Maria; Wirtz, Denis; Tseng, Yiider

    2005-01-28

    MreB, a major component of the bacterial cytoskeleton, exhibits high structural homology to its eukaryotic counterpart actin. Live cell microscopy studies suggest that MreB molecules organize into large filamentous spirals that support the cell membrane and play a key shape-determining function. However, the basic properties of MreB filament assembly remain unknown. Here, we studied the assembly of Thermotoga maritima MreB triggered by ATP in vitro and compared it to the well-studied assembly of actin. These studies show that MreB filament ultrastructure and polymerization depend crucially on temperature as well as the ions present on solution. At the optimal growth temperature of T. maritima, MreB assembly proceeded much faster than that of actin, without nucleation (or nucleation is highly favorable and fast) and with little or no contribution from filament end-to-end annealing. MreB exhibited rates of ATP hydrolysis and phosphate release similar to that of F-actin, however, with a critical concentration of approximately 3 nm, which is approximately 100-fold lower than that of actin. Furthermore, MreB assembled into filamentous bundles that have the ability to spontaneously form ring-like structures without auxiliary proteins. These findings suggest that despite high structural homology, MreB and actin display significantly different assembly properties.

  3. Title: Cytoskeletal proteins in cortical development and diseasesubtitle: Actin associated proteins in periventricular heterotopia

    Directory of Open Access Journals (Sweden)

    Gewei eLian

    2015-04-01

    Full Text Available The actin cytoskeleton regulates many important cellular processes in the brain, including cell division and proliferation, migration, and cytokinesis and differentiation. These developmental processes can be regulated through actin dependent vesicle and organelle movement, cell signaling, and the establishment and maintenance of cell junctions and cell shape. Many of these processes are mediated by extensive and intimate interactions of actin with cellular membranes and proteins. Disruption in the actin cytoskeleton in the brain gives rise to periventricular heterotopia (PH, a malformation of cortical development, characterized by abnormal neurons clustered deep in the brain along the lateral ventricles. This disorder can give rise to seizures, dyslexia and psychiatric disturbances. Anatomically, PH is characterized by a smaller brain (impaired proliferation, heterotopia (impaired initial migration and disruption along the neuroependymal lining (impaired cell-cell adhesion. Genes causal for PH have also been implicated in actin-dependent processes. The current review provides mechanistic insight into actin cytoskeletal regulation of cortical development in the context of this malformation of cortical development.

  4. Release of muscle α-actin into serum after intensive exercise

    Directory of Open Access Journals (Sweden)

    A Martínez-Amat

    2010-12-01

    Full Text Available Purpose: To study the effects of high-level matches on serum alpha actin and other muscle damage markers in teams of rugby and handball players. Methods: Blood samples were drawn from 23 sportsmen: 13 rugby players and 10 handball players. One sample was drawn with the player at rest before the match and one immediately after the match. Immunoassays were used to determine troponin I, troponin T, LDH, and myoglobin concentrations. Western blot and densitometry were used to measure α-actin concentrations. Muscle injury was defined by a total CK value of > 500 IU/L (Rosalki method. Results: Mean pre- and post-match serum alpha-actin values were, respectively, 7.16 and 26.47 μg/ml in the handball group and 1.24 and 20.04 μg/ml in the rugby team. CPK, LDH and myoglobin but not troponin 1 levels also significantly differed between these time points. According to these results, large amounts of α-actin are released into peripheral blood immediately after intense physical effort. Possible cross-interference between skeletal and cardiac muscle damage can be discriminated by the combined use of α-actin and troponin I. Conclusion: The significant increase in alpha-actin after a high-level match may be a reliable marker for the early diagnosis and hence more effective treatment of muscle injury.

  5. Rho-GTPase effector ROCK phosphorylates cofilin in actin-meditated cytokinesis during mouse oocyte meiosis.

    Science.gov (United States)

    Duan, Xing; Liu, Jun; Dai, Xiao-Xin; Liu, Hong-Lin; Cui, Xiang-Shun; Kim, Nam-Hyung; Wang, Zhen-Bo; Wang, Qiang; Sun, Shao-Chen

    2014-02-01

    During oocyte meiosis, a spindle forms in the central cytoplasm and migrates to the cortex. Subsequently, the oocyte extrudes a small body and forms a highly polarized egg; this process is regulated primarily by actin. ROCK is a Rho-GTPase effector that is involved in various cellular functions, such as stress fiber formation, cell migration, tumor cell invasion, and cell motility. In this study, we investigated possible roles for ROCK in mouse oocyte meiosis. ROCK was localized around spindles after germinal vesicle breakdown and was colocalized with cytoplasmic actin and mitochondria. Disrupting ROCK activity by RNAi or an inhibitor resulted in cell cycle progression and polar body extrusion failure. Time-lapse microscopy showed that this may have been due to spindle migration and cytokinesis defects, as chromosomes segregated but failed to extrude a polar body and then realigned. Actin expression at oocyte membranes and in cytoplasm was significantly decreased after these treatments. Actin caps were also disrupted, which was confirmed by a failure to form cortical granule-free domains. The mitochondrial distribution was also disrupted, which indicated that mitochondria were involved in the ROCK-mediated actin assembly. In addition, the phosphorylation levels of Cofilin, a downstream molecule of ROCK, decreased after disrupting ROCK activity. Thus, our results indicated that a ROCK-Cofilin-actin pathway regulated meiotic spindle migration and cytokinesis during mouse oocyte maturation.

  6. Identification of cation-binding sites on actin that drive polymerization and modulate bending stiffness

    Science.gov (United States)

    Kang, Hyeran; Bradley, Michael J.; McCullough, Brannon R.; Pierre, Anaëlle; Grintsevich, Elena E.; Reisler, Emil; De La Cruz, Enrique M.

    2012-01-01

    The assembly of actin monomers into filaments and networks plays vital roles throughout eukaryotic biology, including intracellular transport, cell motility, cell division, determining cellular shape, and providing cells with mechanical strength. The regulation of actin assembly and modulation of filament mechanical properties are critical for proper actin function. It is well established that physiological salt concentrations promote actin assembly and alter the overall bending mechanics of assembled filaments and networks. However, the molecular origins of these salt-dependent effects, particularly if they involve nonspecific ionic strength effects or specific ion-binding interactions, are unknown. Here, we demonstrate that specific cation binding at two discrete sites situated between adjacent subunits along the long-pitch helix drive actin polymerization and determine the filament bending rigidity. We classify the two sites as “polymerization” and “stiffness” sites based on the effects that mutations at the sites have on salt-dependent filament assembly and bending mechanics, respectively. These results establish the existence and location of the cation-binding sites that confer salt dependence to the assembly and mechanics of actin filaments. PMID:23027950

  7. T lymphocyte migration: an action movie starring the actin and associated actors

    Directory of Open Access Journals (Sweden)

    Loïc eDupré

    2015-11-01

    Full Text Available The actin cytoskeleton is composed of a dynamic filament meshwork that builds the architecture of the cell to sustain its fundamental properties. This physical structure is characterized by a continuous remodeling, which allows cells to accomplish complex motility steps such as directed migration, crossing of biological barriers and interaction with other cells. T lymphocytes excel in these motility steps to ensure their immune surveillance duties. In particular, actin cytoskeleton remodeling is key to facilitate the journey of T lymphocytes through distinct tissue environments and to tune their stop and go behavior during the scanning of antigen-presenting cells. The molecular mechanisms controlling actin cytoskeleton remodeling during T lymphocyte motility have been only partially unraveled, since the function of many actin regulators has not yet been assessed in these cells. Our review aims to integrate the current knowledge into a comprehensive picture of how the actin cytoskeleton drives T lymphocyte migration. We will present the molecular actors that control actin cytoskeleton remodeling, as well as their role in the different T lymphocyte motile steps. We will also highlight which challenges remain to be addressed experimentally and which approaches appear promising to tackle them.

  8. T Lymphocyte Migration: An Action Movie Starring the Actin and Associated Actors.

    Science.gov (United States)

    Dupré, Loïc; Houmadi, Raïssa; Tang, Catherine; Rey-Barroso, Javier

    2015-01-01

    The actin cytoskeleton is composed of a dynamic filament meshwork that builds the architecture of the cell to sustain its fundamental properties. This physical structure is characterized by a continuous remodeling, which allows cells to accomplish complex motility steps such as directed migration, crossing of biological barriers, and interaction with other cells. T lymphocytes excel in these motility steps to ensure their immune surveillance duties. In particular, actin cytoskeleton remodeling is a key to facilitate the journey of T lymphocytes through distinct tissue environments and to tune their stop and go behavior during the scanning of antigen-presenting cells. The molecular mechanisms controlling actin cytoskeleton remodeling during T lymphocyte motility have been only partially unraveled, since the function of many actin regulators has not yet been assessed in these cells. Our review aims to integrate the current knowledge into a comprehensive picture of how the actin cytoskeleton drives T lymphocyte migration. We will present the molecular actors that control actin cytoskeleton remodeling, as well as their role in the different T lymphocyte motile steps. We will also highlight which challenges remain to be addressed experimentally and which approaches appear promising to tackle them.

  9. WHAMM links actin assembly via the Arp2/3 complex to autophagy.

    Science.gov (United States)

    Kast, David J; Dominguez, Roberto

    2015-01-01

    Macroautophagy (hereafter autophagy) is the process by which cytosolic material destined for degradation is enclosed inside a double-membrane cisterna known as the autophagosome and processed for secretion and/or recycling. This process requires a large collection of proteins that converge on certain sites of the ER membrane to generate the autophagosome membrane. Recently, it was shown that actin accumulates around autophagosome precursors and could play a role in this process, but the mechanism and role of actin polymerization in autophagy were unknown. Here, we discuss our recent finding that the nucleation-promoting factor (NPF) WHAMM recruits and activates the Arp2/3 complex for actin assembly at sites of autophagosome formation on the ER. Using high-resolution, live-cell imaging, we showed that WHAMM forms dynamic puncta on the ER that comigrate with several autophagy markers, and propels the spiral movement of these puncta by an Arp2/3 complex-dependent actin comet tail mechanism. In starved cells, WHAMM accumulates at the interface between neighboring autophagosomes, whose number and size increases with WHAMM expression. Conversely, knocking down WHAMM, inhibiting the Arp2/3 complex or interfering with actin polymerization reduces the size and number of autophagosomes. These findings establish a link between Arp2/3 complex-mediated actin assembly and autophagy.

  10. Displacement of the mitotic apparatuses by centrifugation reveals cortical actin organization during cytokinesis in cultured tobacco BY-2 cells.

    Science.gov (United States)

    Arima, Kengo; Tamaoki, Daisuke; Mineyuki, Yoshinobu; Yasuhara, Hiroki; Nakai, Tomonori; Shimmen, Teruo; Yoshihisa, Tohru; Sonobe, Seiji

    2018-06-19

    In plant cytokinesis, actin is thought to be crucial in cell plate guidance to the cortical division zone (CDZ), but its organization and function are not fully understood. To elucidate actin organization during cytokinesis, we employed an experimental system, in which the mitotic apparatus is displaced and separated from the CDZ by centrifugation and observed using a global-local live imaging microscope that enabled us to record behavior of actin filaments in the CDZ and the whole cell division process in parallel. In this system, returning movement of the cytokinetic apparatus in cultured-tobacco BY-2 cells occurs, and there is an advantage to observe actin organization clearly during the cytokinetic phase because more space was available between the CDZ and the distantly formed phragmoplast. Actin cables were clearly observed between the CDZ and the phragmoplast in BY-2 cells expressing GFP-fimbrin after centrifugation. Both the CDZ and the edge of the expanding phragmoplast had actin bulges. Using live-cell imaging including the global-local live imaging microscopy, we found actin filaments started to accumulate at the actin-depleted zone when cell plate expansion started even in the cell whose cell plate failed to reach the CDZ. These results suggest that specific accumulation of actin filaments at the CDZ and the appearance of actin cables between the CDZ and the phragmoplast during cell plate formation play important roles in the guidance of cell plate edges to the CDZ.

  11. Azadirachtin(A) distinctively modulates subdomain 2 of actin - novel mechanism to induce depolymerization revealed by molecular dynamics study.

    Science.gov (United States)

    Pravin Kumar, R; Roopa, L; Sudheer Mohammed, M M; Kulkarni, Naveen

    2016-12-01

    Azadirachtin(A) (AZA), a potential insecticide from neem, binds to actin and induces depolymerization in Drosophila. AZA binds to the pocket same as that of Latrunculin A (LAT), but LAT inhibits actin polymerization by stiffening the actin structure and affects the ADP-ATP exchange. The mechanism by which AZA induces actin depolymerization is not clearly understood. Therefore, different computational experiments were conducted to delineate the precise mechanism of AZA-induced actin depolymerization. Molecular dynamics studies showed that AZA strongly interacted with subdomain 2 and destabilized the interactions between subdomain 2 of one actin and subdomains 1 and 4 of the adjacent actin, causing the separation of actin subunits. The separation was observed between subdomain 3 of subunit n and subdomain 4 of subunit n + 2. However, the specific triggering point for the separation of the subunits was the destabilization of direct interactions between subdomain 2 of subunit n (Arg39, Val45, Gly46 and Arg62) and subdomain 4 of subunit n + 2 (Asp286, Ile287, Asp288, Ile289, Asp244 and Lys291). These results reveal a unique mechanism of an actin filament modulator that induces depolymerization. This mechanism of AZA can be used to design similar molecules against mammalian actins for cancer therapy.

  12. FIMBRIN1 Is Involved in Lily Pollen Tube Growth by Stabilizing the Actin Fringe[C][W][OA

    Science.gov (United States)

    Su, Hui; Zhu, Jinsheng; Cai, Chao; Pei, Weike; Wang, Jiaojiao; Dong, Huaijian; Ren, Haiyun

    2012-01-01

    An actin fringe structure in the subapex plays an important role in pollen tube tip growth. However, the precise mechanism by which the actin fringe is generated and maintained remains largely unknown. Here, we cloned a 2606-bp full-length cDNA encoding a deduced 77-kD fimbrin-like protein from lily (Lilium longiflorum), named FIMBRIN1 (FIM1). Ll-FIM1 was preferentially expressed in pollen and concentrated at actin fringe in the subapical region, as well as in longitudinal actin-filament bundles in the shank of pollen tubes. Microinjection of Ll-FIM1 antibody into lily pollen tubes inhibited tip growth and disrupted the actin fringe. Furthermore, we verified the function of Ll-FIM1 in the fim5 mutant of its closest relative, Arabidopsis thaliana. Pollen tubes of fim5 mutants grew with a larger diameter in early stages but could recover into normal forms in later stages, despite significantly slower growth rates. The actin fringe of the fim5 mutants, however, was impaired during both early and late stages. Impressively, stable expression of fim5pro:GFP:Ll-FIM1 rescued the actin fringe and the growth rate of Arabidopsis fim5 pollen tubes. In vitro biochemical analysis showed that Ll-FIM1 could bundle actin filaments. Thus, our study has identified a fimbrin that may stabilize the actin fringe by cross-linking actin filaments into bundles, which is important for proper tip growth of lily pollen tubes. PMID:23150633

  13. G-actin sequestering protein thymosin-β4 regulates the activity of myocardin-related transcription factor

    International Nuclear Information System (INIS)

    Morita, Tsuyoshi; Hayashi, Ken’ichiro

    2013-01-01

    Highlights: •Tβ4 competed with MRTF-A for G-actin binding. •Tβ4 activated the MRTF–SRF signaling pathway. •Tβ4 increased the endogenous expression of SRF-dependent genes. -- Abstract: Myocardin-related transcription factors (MRTFs) are robust coactivators of serum response factor (SRF). MRTFs contain three copies of the RPEL motif at their N-terminus, and they bind to monomeric globular actin (G-actin). Previous studies illustrate that G-actin binding inhibits MRTF activity by preventing the MRTFs nuclear accumulation. In the living cells, the majority of G-actin is sequestered by G-actin binding proteins that prevent spontaneous actin polymerization. Here, we demonstrate that the most abundant G-actin sequestering protein thymosin-β4 (Tβ4) was involved in the regulation of subcellular localization and activity of MRTF-A. Tβ4 competed with MRTF-A for G-actin binding; thus, interfering with G-actin–MRTF-A complex formation. Tβ4 overexpression induced the MRTF-A nuclear accumulation and activation of MRTF–SRF signaling. The activation rate of MRTF-A by the Tβ4 mutant L17A, whose affinity for G-actin is very low, was lower than that by wild-type Tβ4. In contrast, the β-actin mutant 3DA, which has a lower affinity for Tβ4, more effectively suppressed MRTF-A activity than wild-type β-actin. Furthermore, ectopic Tβ4 increased the endogenous expression of SRF-dependent actin cytoskeletal genes. Thus, Tβ4 is an important MRTF regulator that controls the G-actin–MRTFs interaction

  14. G-actin sequestering protein thymosin-β4 regulates the activity of myocardin-related transcription factor

    Energy Technology Data Exchange (ETDEWEB)

    Morita, Tsuyoshi, E-mail: tsuyo@nbiochem.med.osaka-u.ac.jp; Hayashi, Ken’ichiro

    2013-08-02

    Highlights: •Tβ4 competed with MRTF-A for G-actin binding. •Tβ4 activated the MRTF–SRF signaling pathway. •Tβ4 increased the endogenous expression of SRF-dependent genes. -- Abstract: Myocardin-related transcription factors (MRTFs) are robust coactivators of serum response factor (SRF). MRTFs contain three copies of the RPEL motif at their N-terminus, and they bind to monomeric globular actin (G-actin). Previous studies illustrate that G-actin binding inhibits MRTF activity by preventing the MRTFs nuclear accumulation. In the living cells, the majority of G-actin is sequestered by G-actin binding proteins that prevent spontaneous actin polymerization. Here, we demonstrate that the most abundant G-actin sequestering protein thymosin-β4 (Tβ4) was involved in the regulation of subcellular localization and activity of MRTF-A. Tβ4 competed with MRTF-A for G-actin binding; thus, interfering with G-actin–MRTF-A complex formation. Tβ4 overexpression induced the MRTF-A nuclear accumulation and activation of MRTF–SRF signaling. The activation rate of MRTF-A by the Tβ4 mutant L17A, whose affinity for G-actin is very low, was lower than that by wild-type Tβ4. In contrast, the β-actin mutant 3DA, which has a lower affinity for Tβ4, more effectively suppressed MRTF-A activity than wild-type β-actin. Furthermore, ectopic Tβ4 increased the endogenous expression of SRF-dependent actin cytoskeletal genes. Thus, Tβ4 is an important MRTF regulator that controls the G-actin–MRTFs interaction.

  15. The F-Actin Binding Protein Cortactin Regulates the Dynamics of the Exocytotic Fusion Pore through its SH3 Domain

    Science.gov (United States)

    González-Jamett, Arlek M.; Guerra, María J.; Olivares, María J.; Haro-Acuña, Valentina; Baéz-Matus, Ximena; Vásquez-Navarrete, Jacqueline; Momboisse, Fanny; Martinez-Quiles, Narcisa; Cárdenas, Ana M.

    2017-01-01

    Upon cell stimulation, the network of cortical actin filaments is rearranged to facilitate the neurosecretory process. This actin rearrangement includes both disruption of the preexisting actin network and de novo actin polymerization. However, the mechanism by which a Ca2+ signal elicits the formation of new actin filaments remains uncertain. Cortactin, an actin-binding protein that promotes actin polymerization in synergy with the nucleation promoting factor N-WASP, could play a key role in this mechanism. We addressed this hypothesis by analyzing de novo actin polymerization and exocytosis in bovine adrenal chromaffin cells expressing different cortactin or N-WASP domains, or cortactin mutants that fail to interact with proline-rich domain (PRD)-containing proteins, including N-WASP, or to be phosphorylated by Ca2+-dependent kinases, such as ERK1/2 and Src. Our results show that the activation of nicotinic receptors in chromaffin cells promotes cortactin translocation to the cell cortex, where it colocalizes with actin filaments. We further found that, in association with PRD-containing proteins, cortactin contributes to the Ca2+-dependent formation of F-actin, and regulates fusion pore dynamics and the number of exocytotic events induced by activation of nicotinic receptors. However, whereas the actions of cortactin on the fusion pore dynamics seems to depend on the availability of monomeric actin and its phosphorylation by ERK1/2 and Src kinases, cortactin regulates the extent of exocytosis by a mechanism independent of actin polymerization. Together our findings point out a role for cortactin as a critical modulator of actin filament formation and exocytosis in neuroendocrine cells. PMID:28522963

  16. Characterization of the activities of actin-affecting drugs on tumor cell migration

    International Nuclear Information System (INIS)

    Hayot, Caroline; Debeir, Olivier; Ham, Philippe van; Damme, Marc van; Kiss, Robert; Decaestecker, Christine

    2006-01-01

    Metastases kill 90% of cancer patients. It is thus a major challenge in cancer therapy to inhibit the spreading of tumor cells from primary tumor sites to those particular organs where metastases are likely to occur. Whereas the actin cytoskeleton is a key component involved in cell migration, agents targeting actin dynamics have been relatively poorly investigated. Consequently, valuable in vitro pharmacological tools are needed to selectively identify this type of agent. In response to the absence of any standardized process, the present work aims to develop a multi-assay strategy for screening actin-affecting drugs with anti-migratory potentials. To validate our approach, we used two cancer cell lines (MCF7 and A549) and three actin-affecting drugs (cytochalasin D, latrunculin A, and jasplakinolide). We quantified the effects of these drugs on the kinetics of actin polymerization in tubes (by means of spectrofluorimetry) and on the dynamics of actin cytoskeletons within whole cells (by means of fluorescence microscopy). Using quantitative videomicroscopy, we investigated the actual effects of the drugs on cell motility. Finally, the combined drug effects on cell motility and cell growth were evaluated by means of a scratch-wound assay. While our results showed concordant drug-induced effects on actin polymerization occurring in vitro in test tubes and within whole cells, the whole cell assay appeared more sensitive than the tube assay. The inhibition of actin polymerization induced by cytochalasin D was paralleled by a decrease in cell motility for both cell types. In the case of jasplakinolide, which induces actin polymerization, while it significantly enhanced the locomotion of the A549 cells, it significantly inhibited that of the MCF-7 ones. All these effects were confirmed by means of the scratch-wound assay except of the jasplakinolide-induced effects on MCF-7 cell motility. These later seemed compensated by an additional effect occurring during wound

  17. Dual-channel red/blue fluorescence dosimetry with broadband reflectance spectroscopic correction measures protoporphyrin IX production during photodynamic therapy of actinic keratosis

    Science.gov (United States)

    Kanick, Stephen Chad; Davis, Scott C.; Zhao, Yan; Hasan, Tayyaba; Maytin, Edward V.; Pogue, Brian W.; Chapman, M. Shane

    2014-07-01

    Dosimetry for aminolevulinic acid (ALA)-induced protoporphyrin IX (PpIX) photodynamic therapy of actinic keratosis was examined with an optimized fluorescence dosimeter to measure PpIX during treatment. While insufficient PpIX generation may be an indicator of incomplete response, there exists no standardized method to quantitate PpIX production at depths in the skin during clinical treatments. In this study, a spectrometer-based point probe dosimeter system was used to sample PpIX fluorescence from superficial (blue wavelength excitation) and deeper (red wavelength excitation) tissue layers. Broadband white light spectroscopy (WLS) was used to monitor aspects of vascular physiology and inform a correction of fluorescence for the background optical properties. Measurements in tissue phantoms showed accurate recovery of blood volume fraction and reduced scattering coefficient from WLS, and a linear response of PpIX fluorescence versus concentration down to 1.95 and 250 nM for blue and red excitations, respectively. A pilot clinical study of 19 patients receiving 1-h ALA incubation before treatment showed high intrinsic variance in PpIX fluorescence with a standard deviation/mean ratio of >0.9. PpIX fluorescence was significantly higher in patients reporting higher pain levels on a visual analog scale. These pilot data suggest that patient-specific PpIX quantitation may predict outcome response.

  18. Major therapeutic effect of pentoxifylline-tocopherol association in the superficial radioinduced fibrosis: phase II test

    International Nuclear Information System (INIS)

    Delanian, S.; Balla-Mekias, S.; Maylin, C.; Lefaix, J.L.

    1997-01-01

    The association of pentoxifylline-tocopherol seems efficient in the reduction of the superficial human radioinduced fibrosis. This phase II invites to realize a randomized test and to a comparison with the results got with the dismutase superoxide. (N.C.)

  19. Primary temporal region squamous cell carcinoma diagnosed by a superficial temporal artery biopsy

    DEFF Research Database (Denmark)

    Andersen, S A W; Kiss, K

    2015-01-01

    artery biopsy was performed. The histopathology revealed perineural invasion of squamous cell carcinoma (SCC). A thorough investigation revealed no other primary site for the SCC and the patient was treated with surgical excision. CONCLUSION: Malignancy is rarely found in superficial temporal artery...

  20. Automatic and Deliberate Affective Associations with Sexual Stimuli in Women with Superficial Dyspareunia

    NARCIS (Netherlands)

    Brauer, Marieke; de Jong, Peter J.; Huijding, Jorg; Laan, Ellen; ter Kuile, Moniek M.

    Current views suggest that in women with superficial dyspareunia the prospect of penile-vaginal intercourse automatically activates fear-related associations. The automatic activation of negative associations is assumed to interfere with the development of sexual arousal. In turn, this may further

  1. Treatment of primary cutaneous anaplastic large cell lymphoma with superficial x-rays

    DEFF Research Database (Denmark)

    Jepsen, Malene E; Gniadecki, Robert

    2015-01-01

    The optimal radiation schedule for primary cutaneous anaplastic lymphoma (PCALCL) has not been investigated. We report here satisfactory outcomes of low-dose (16-20 Gy, 3-5 fractions), superficial X-ray radiation (40-50 kV) in a series of 10 patients with PCALCL. Only 1 patient developed a local...

  2. Removal of dissolved organic carbon in pilot wetlands of subsuperficial and superficial flows

    Directory of Open Access Journals (Sweden)

    Ruth M. Agudelo C

    2010-04-01

    Full Text Available Objective: to compare removal of dissolved organic carbon (d o c obtained with pilot wetlands of subsuperficial flow (p h s s and superficial flow (p h s, with Phragmites australis as treatment alternatives for domestic residual waters of small communities and rural areas. Methodology: an exploratory and experimental study was carried out adding 100,12 mg/L of dissolved organic carbon to synthetic water contaminated with Chlorpyrifos in order to feed the wetlands. A total amount of 20 samples were done, 16 of them in four experiments and the other ones in the intervals with no use of pesticides. Samples were taken on days 1, 4, 8, and 11 in the six wetlands, three of them subsuperficial, and three of them superficial. The main variable answer was dissolved organic carbon, measured in the organic carbon analyzer. Results: a high efficiency in the removal of d o c was obtained with the two types of wetlands: 92,3% with subsuperficial flow and 95,6% with superficial flow. Such a high removal was due to the interaction between plants, gravel and microorganisms. Conclusion: although in both types of wetlands the removal was high and similar, it is recommended to use those of subsuperficial flow because in the superficial ones algae and gelatinous bio-films are developed, which becomes favorable to the development of important epidemiologic vectors in terms of public health.

  3. Honey compared with silver sulphadiazine in the treatment of superficial partial-thickness burns.

    Science.gov (United States)

    Malik, Kamran Ishaque; Malik, M A Nasir; Aslam, Azhar

    2010-10-01

    Burn injury is associated with a high incidence of death and disability; yet, its management remains problematic and costly. We conducted this clinical study to evaluate the efficacy of honey in the treatment of superficial and partial-thickness burns covering less than 40% of body surface area and compared its results with those of silver sulphadiazine (SSD). In this randomised comparative clinical trial, carried out Burn Center of POF Hospital, Wah Cantt, Pakistan, from May 2007 to February 2008, 150 patients of all ages having similar types of superficial and partial-thickness burns at two sites on different parts of body were included. Each patient had one burn site treated with honey and one treated with topical SSD, randomly. The rate of re-epithelialization and healing of superficial and partial-thickness burns was significantly faster in the sites treated with honey than in the sites treated with SSD (13·47 ± 4·06 versus 15·62 ± 4·40 days, respectively: P honey healed completely in less than 21 days versus 24 days for the site treated with SSD. Six patients had positive culture for Pseudomonas aeroginsa in honey-treated site, whereas 27 patients had positive culture in SSD-treated site. The results clearly showed greater efficacy of honey over SSD cream for treating superficial and partial-thickness burns. 2010 The Authors. Journal Compilation © 2010 Blackwell Publishing Ltd and Medicalhelplines.com Inc.

  4. Thrombophlebitis of the penile superficial vein, penile mondor's disease: a case report

    Energy Technology Data Exchange (ETDEWEB)

    Park, Sang Hyun; Kim, Young Hwa; Kim, Doo Sang; Shin, Hyung Chul; Bae, Won Kyung; Kim, Il Young [Soonchunhyang University, Chunan (Korea, Republic of); Kim, Hyun Cheol [Kyung-Hee University, Seoul (Korea, Republic of)

    2007-07-15

    Mondor's disease is commonly known as thrombophlebitis of the superficial vein in the breast, and this disease occurs rarely in the penis. Despite extensive information about the clinical presentation and course of this disease, imaging findings for this disease are limited. We report gray scale and power Doppler sonographic findings of penile Mondor's disease.

  5. Protocolo nacional de monitoreo de la calidad de los en cuerpos naturales de agua superficial

    OpenAIRE

    Autoridad Nacional del Agua. Dirección de Gestión de Calidad de los Recursos Hídricos

    2011-01-01

    Estandariza la metodología para el desarrollo del monitoreo de la calidad de los recursos hídricos en los cuerpos naturales de agua superficial, asimismo pretende convertirse en un instrumento de gestión en el desarrollo del Plan Nacional de Vigilancia de la Calidad de Agua en el Perú.

  6. The defence architecture of the superficial cells of the oral mucosa

    NARCIS (Netherlands)

    Asikainen, P.; Ruotsalainen, T.J.; Mikkonen, J.J.W.; Koistinen, A.; ten Bruggenkate, C.M.; Kullaa, A.M.

    2012-01-01

    The oral epithelium together with the saliva and its components forms a complex structure which is the first line of defence in the oral cavity. The surface of superficial cells of the oral epithelium contains ridge-like folds, microplicae (MPL), which are typical of the surfaces of areas covered

  7. The Effects of Test Anxiety on Learning at Superficial and Deep Levels of Processing.

    Science.gov (United States)

    Weinstein, Claire E.; And Others

    1982-01-01

    Using a deep-level processing strategy, low test-anxious college students performed significantly better than high test-anxious students in learning a paired-associate word list. Using a superficial-level processing strategy resulted in no significant difference in performance. A cognitive-attentional theory and test anxiety mechanisms are…

  8. The flexural stiffness of superficial neuromasts in the zebrafish (Danio rerio) lateral line

    NARCIS (Netherlands)

    McHenry, Matthew J.; van Netten, Sietse M.

    2007-01-01

    Superficial neuromasts are structures that detect water flow on the surface of the body of fish and amphibians. As a component of the lateral line system, these receptors are distributed along the body, where they sense flow patterns that mediate a wide variety of behaviors. Their ability to detect

  9. Subintimal angioplasty for the treatment of long segment occlusion of superficial femoral artery: the midterm results

    International Nuclear Information System (INIS)

    Lou Wensheng; Gu Jianping; He Xu; Chen Liang; Chen Guoping; Su Haobo; Song Jinhua; Wang Tao; Xu Ke

    2011-01-01

    Objective: To discuss the clinical value of subintimal angioplasty in treating long segment occlusion of superficial femoral artery and to observe its midterm results. Methods: Subintimal angioplasty was performed in 45 patients with long segment occlusion of superficial femoral artery, whose clinical presentation was intermittent claudication or critical limb ischemia. The primary patency, limb salvage and factors influencing long-term patency were observed, and the clinical data were analyzed. Results: Of the total 45 cases, the subintimal angioplasty was successfully accomplished in 43. The success rate of antegrade approach technique via the superficial femoral artery was 80% (n=36), the occlusion was recanalized by using retrograde approach technique via ipsilateral popliteal artery in 7 case. The total technical success rate was 95.6%. The primary patency in 43 successful cases at 6, 12, 24 and 36 months was 85.7%, 69.0% 57.3% and 50.9%, respectively. No statistically significant difference in primary patency existed between the group of intermittent claudication and the group of critical limb ischemia (P>0.05). Statistically significant prediction factors for primary patency included the number of the run off vessels below the knee and the length of the occlusion (P<0.05). Conclusion: Subintimal angioplasty is an effective procedure for the treatment of long segment occlusions of superficial femoral artery with satisfactory mid-term results. (authors)

  10. The analogy between dreams and the ancient art of memory is tempting but superficial.

    Science.gov (United States)

    Axmacher, Nikolai; Fell, Juergen

    2013-12-01

    Although the analogy between dreams and ancient mnemotechniques is tempting because they share several phenomenological characteristics, this analogy is superficial at a closer look. Unlike mneomotechnically encoded material, rapid eye movement (REM) dreams are inherently difficult to remember, do not usually allow conscious subsequent retrieval of all interconnected elements, and have been found to support subsequent episodic memory in only rare cases.

  11. Masaje circulatorio superficial en la enfermedad arterial periférica

    OpenAIRE

    Cebrià Iranzo, M. Àngels

    2010-01-01

    En la enfermedad arterial periférica crónica se indica el masaje circulatorio superficial por producir una vasodilatación refleja en las zonas afectadas. Este masaje manual tiene dos objetivos fundamentales: 1) Favorecer la vasodilatación refleja e hiperemia local; 2) Drenar el retorno vascular sanguíneo y linfático.

  12. Percutaneous transluminal angioplasty of the superficial femoral artery by retrograde catheterization via the popliteal artery

    DEFF Research Database (Denmark)

    Tønnesen, K H; Sager, P; Karle, A

    1988-01-01

    We report the results of 50 angioplasty procedures via the popliteal artery. A 3-year follow-up including control of blood pressures at ankle and toe levels show results comparable to reports in the literature. This new approach for angioplasty of the superficial femoral artery and eventually of ...

  13. Induction of superficial cortical layer neurons from mouse embryonic stem cells by valproic acid.

    Science.gov (United States)

    Juliandi, Berry; Abematsu, Masahiko; Sanosaka, Tsukasa; Tsujimura, Keita; Smith, Austin; Nakashima, Kinichi

    2012-01-01

    Within the developing mammalian cortex, neural progenitors first generate deep-layer neurons and subsequently more superficial-layer neurons, in an inside-out manner. It has been reported recently that mouse embryonic stem cells (mESCs) can, to some extent, recapitulate cortical development in vitro, with the sequential appearance of neurogenesis markers resembling that in the developing cortex. However, mESCs can only recapitulate early corticogenesis; superficial-layer neurons, which are normally produced in later developmental periods in vivo, are under-represented. This failure of mESCs to reproduce later corticogenesis in vitro implies the existence of crucial factor(s) that are absent or uninduced in existing culture systems. Here we show that mESCs can give rise to superficial-layer neurons efficiently when treated with valproic acid (VPA), a histone deacetylase inhibitor. VPA treatment increased the production of Cux1-positive superficial-layer neurons, and decreased that of Ctip2-positive deep-layer neurons. These results shed new light on the mechanisms of later corticogenesis. Copyright © 2011 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

  14. Improved superficial brain hemorrhage visualization in susceptibility weighted images by constrained minimum intensity projection

    Science.gov (United States)

    Castro, Marcelo A.; Pham, Dzung L.; Butman, John

    2016-03-01

    Minimum intensity projection is a technique commonly used to display magnetic resonance susceptibility weighted images, allowing the observer to better visualize hemorrhages and vasculature. The technique displays the minimum intensity in a given projection within a thick slab, allowing different connectivity patterns to be easily revealed. Unfortunately, the low signal intensity of the skull within the thick slab can mask superficial tissues near the skull base and other regions. Because superficial microhemorrhages are a common feature of traumatic brain injury, this effect limits the ability to proper diagnose and follow up patients. In order to overcome this limitation, we developed a method to allow minimum intensity projection to properly display superficial tissues adjacent to the skull. Our approach is based on two brain masks, the largest of which includes extracerebral voxels. The analysis of the rind within both masks containing the actual brain boundary allows reclassification of those voxels initially missed in the smaller mask. Morphological operations are applied to guarantee accuracy and topological correctness, and the mean intensity within the mask is assigned to all outer voxels. This prevents bone from dominating superficial regions in the projection, enabling superior visualization of cortical hemorrhages and vessels.

  15. Effect of exposing pupae of flesh fly Parasarcophaga ruficornis F. to superficial X-rays

    International Nuclear Information System (INIS)

    Kumar, K.; Srivastava, U.S.

    1991-01-01

    Irradiation of pupae of Parasarcophaga ruficornis with different doses of superficial X-rays leads to lethality and interference with moulting and metamorphosis. Younger pupae are more radiosensitive in respect of lethality and metamorphosis to older ones. (author). 12 ref., 9 figs., 1 tab

  16. Dorsal stress fibers, transverse actin arcs, and perinuclear actin fibers form an interconnected network that induces nuclear movement in polarizing fibroblasts

    Czech Academy of Sciences Publication Activity Database

    Maninová, Miloslava; Vomastek, Tomáš

    2016-01-01

    Roč. 283, č. 20 (2016), s. 3676-3693 ISSN 1742-464X R&D Projects: GA ČR GA13-06405S Institutional support: RVO:61388971 Keywords : actin dorsal fibers * cell polarity * nuclear reorientation Subject RIV: EE - Microbiology, Virology Impact factor: 3.902, year: 2016

  17. Defining the actinic keratosis field: a literature review and discussion.

    Science.gov (United States)

    Figueras Nart, I; Cerio, R; Dirschka, T; Dréno, B; Lear, J T; Pellacani, G; Peris, K; Ruiz de Casas, A

    2018-04-01

    Despite the chronic and increasingly prevalent nature of actinic keratosis (AK) and existing evidence supporting assessment of the entire cancerization field during clinical management, a standardized definition of the AK field to aid in the understanding and characterization of the disease is lacking. The objective of this review was to present and appraise the available evidence describing the AK cancerization field, with the aim of determining a precise definition of the AK field in terms of its molecular (including genetic and immunological), histological and clinical characteristics. Eight European dermatologists collaborated to conduct a review and expert appraisal of articles detailing the characteristics of the AK field. Articles published in English before August 2016 were identified using PubMed and independently selected for further assessment according to predefined preliminary inclusion and exclusion criteria. In addition, a retrospective audit of patients with AK was performed to define the AK field in clinical terms. A total of 32 review articles and 47 original research articles provided evidence of sun-induced molecular (including genetic and immunological) and histological skin changes in the sun-exposed area affected by AK. However, the available literature was deemed insufficient to inform a clinical definition of the AK field. During the retrospective audit, visible signs of sun damage in 40 patients with AK were assessed. Telangiectasia, atrophy and pigmentation disorders emerged as 'reliable or very reliable' indicators of AK field based on expert opinion, whereas 'sand paper' was deemed a 'moderately reliable' indicator. This literature review has revealed a significant gap of evidence to inform a clinical definition of the AK field. Therefore, the authors instead propose a clinical definition of field cancerization based on the identification of visible signs of sun damage that are reliable indicators of field cancerization based on expert

  18. Prevalence of actinic keratosis among dermatology outpatients in Spain.

    Science.gov (United States)

    Ferrándiz, C; Plazas, M J; Sabaté, M; Palomino, R

    2016-10-01

    Actinic keratoses (AKs) are common skin lesions associated with an increased risk of developing squamous cell carcinoma. Few studies in Europe have focused on AK prevalence. To determine the point prevalence of AKs in a dermatology outpatient population in Spain, to describe the clinical characteristics of these lesions and to characterise the profile of AK patients. Observational, cross-sectional, multicentre study conducted in 19 hospitals (dermatology outpatient services) around Spain. A total of 204 consecutive patients per hospital who were ≥45 years old were screened for the presence of AKs. 3877 patients were assessed and the overall AKs prevalence was 28.6%. Prevalence was significantly higher in men than women (38.4% vs. 20.8%, p<0.0001) and increased with age for both sexes (45.2% in 71-80 years). Scalp and ear lesion locations were significantly more frequent in men (51.9% vs. 2.7% and 16.9% vs. 2.4%, respectively, p<0.0001 both cases) and the cheek, nose and neckline in women (46.3% vs. 34.0% [p<0.0001], 43.0% vs. 24.8% [p<0.0001] and 5.3% vs. 1.8% [p=0.002]). Men showed a significantly higher frequency of ≥2 affected areas than women (42.7% vs. 20.3%, p<0.0001). Among patients with AK lesions, only 65% confirmed that they were the reason for the visit to the clinic. Approximately a quarter of the dermatology outpatient population in Spain aged ≥45 years old have AKs, with the prevalence rate being highest in men and in older age groups. AK is underdiagnosed and a proactive strategy is needed for the diagnosis and early treatment of these lesions. Copyright © 2016 AEDV. Publicado por Elsevier España, S.L.U. All rights reserved.

  19. Pneumodissection for skin protection in image-guided cryoablation of superficial musculoskeletal tumours.

    Science.gov (United States)

    Maybody, Majid; Tang, Peter Q; Moskowitz, Chaya S; Hsu, Meier; Yarmohammadi, Hooman; Boas, F Edward

    2017-03-01

    Pneumodissection is described as a simple method for preventing skin injury during cryoablation of superficial musculoskeletal tumours. Superficial tumour cryoablations performed from 2009 to 2015 were retrospectively reviewed. Pneumodissection was performed in 13 patients when the shortest tumour-skin distance was less than 25 mm. Indications were pain palliation (n = 9) and local tumour control (n = 4). Patients, target tumours, technical characteristics and complications up to 60 days post ablation were reviewed. The ice ball-skin distances with and without pneumodissection were compared by a paired t-test and further assessed for association with covariates using ANCOVA. Technical success for ablation was 12 of 13. The mean shortest tumour-skin distance was 15.0 mm (3.2-24.5 mm). The mean thickness of pneumodissection was 9.6 mm (5.2-16.6 mm) resulting in mean elevation of skin of 3.4 mm (1.2-5.3 mm). Mean shortest ice ball-skin distance after pneumodissection was 10.5 mm (4.2-19.7 mm). No infection or systemic air embolism was noted. No intraprocedural frostbite was observed. Pneumodissection is feasible, effective and safe in protecting the skin during image-guided cryoablation of superficial tumours. • Frostbite during image-guided cryoablation of superficial tumours is commonly under-reported. • Frostbites are painful and may introduce infection into the superficial ablation zone. • Warm compress, saline and CO 2 have shortcomings in protecting the skin. • Pneumodissection is free, readily available, easy to use and safe and effective.

  20. Investigation of the behaviour of a LILW superficial repository under aircraft impact

    International Nuclear Information System (INIS)

    Lo Frano, Rosa; Stefanini, Lorenzo

    2016-01-01

    Highlights: • Safety assessment of a LILW superficial repository. • Investigation of the consequences of an aircraft impact with fuel burning. • Experimental material properties. • Numerical simulation of aircraft impact with fuel burning accident by MSC.MARC"© code. • Demonstration that the overall integrity resulted is guaranteed. - Abstract: Safety and security are the two fundamental aspects to guarantee when designing a LILW superficial repository. Because of its safety concern, we have to prove, and build confidence in, the primary and secondary consequences of the crashing will be acceptable. These goals are obtained generally by means of safety assessment supported by calculations. This study is intended to investigate the performance of a superficial repository subjected to aircraft impact and fuel burning. To the purpose a superficial repository similar to that of El Cabril has been considered. Moreover to be confident the facility is safe and that the consequences of such a type of accident on the environment and humans are negligible, an appropriate safety assessment was carried out. The potential damage that aircraft impact could bring into the repository has been therefore analysed and discussed. To attain the intent load functions, calculated according to the Riera approach, and the maximum temperature reached by fuel during its combustion have been considered. FEM (thermo-mechanical) simulations have been done, by MSC"© Marc code, assuming damaging phenomena of concrete and material properties variation with the temperature. The obtained results showed that an empty superficial repository with a wall thickness, ranging from 0.7 to 1 m, is not sufficient to avoid penetration. Nevertheless even in presence of a reduced strength and of (cone) cracking and plugging, the overall integrity resulted guaranteed.