Distribution by Scientific Domains
Distribution within Medical Sciences

Selected Abstracts

From anchorage dependent proliferation to survival: Lessons from redox signalling

IUBMB LIFE, Issue 5 2008
Paola Chiarugi
Abstract Anchorage to extracellular matrix (ECM) is essential for the execution of the mitotic program of nontransformed cells as they need simultaneous signals starting from mitogenic molecules, as growth factors (GFs), and adhesive agents belonging to ECM. Reactive oxygen species play a key function during both GF and integrin receptor signalling and are therefore recognised to have a synergistic function with several others transducers for anchorage-dependent growth (ADG). Indeed, redox-regulated proteins include protein tyrosine phosphatases, protein tyrosine kinases, small GTPases, cytoskeleton proteins, as well as several transcription factors. In this review, we focus on the role of reactive oxygen species (ROS) as key second messengers granting a proper executed mitosis for anchorage-dependent cells through redox regulation of several downstream targets. Besides, redox signals elicited by ECM contact assure a protection from anoikis, a specific apoptosis induced by lack of anchorage. Cancer cells frequently show a deregulation of ROS production and a constitutive oxidative stress has been associated to the achievement of an invasive phenotype. Hence, in cancer cells, the constitutive deregulation of both mitogenic and survival pathways, likely mimicking autocrine/adhesive signals, helps to guide the transformed cells to escape the innate apoptotic response to abolish the signals started by cell/ECM contact, thus sustaining the spreading of anchorage-independent cancer cells and the metastases growth. © 2008 IUBMB IUBMB Life, 60(5): 301,307, 2008 [source]

Palatal Anchorage for the Retention of Interim Removable Prostheses

Alberto J. Ambard DDS
Abstract This paper describes a technique that involves the use of palatal implants to retain a maxillary interim prosthesis when extensive bone graft procedures are performed. The rationale is that some bone graft procedures require the removal of the denture flanges for graft success. Once the denture flanges are removed, the denture loses all its retention capabilities, making this lengthy interim phase difficult for the patient. While the use of palatal implants has been documented extensively, limited information is available to describe the use of palatal implants for prosthetic reasons. [source]

Corrosion Books: Anchorage in Concrete Construction.

By: Rolf Eligehausen, John F. Silva, Rainer Mallee

Anchorage to the cytosolic face of the endoplasmic reticulum membrane: a new strategy to stabilize a cytosolic recombinant antigen in plants

Alessandra Barbante
Summary The levels of accumulation of recombinant vaccines in transgenic plants are protein specific and strongly influenced by the subcellular compartment of destination. The human immunodeficiency virus protein Nef (negative factor), a promising target for the development of an antiviral vaccine, is a cytosolic protein that accumulates to low levels in transgenic tobacco and is even more unstable when introduced into the secretory pathway, probably because of folding defects in the non-cytosolic environment. To improve Nef accumulation, a new strategy was developed to anchor the molecule to the cytosolic face of the endoplasmic reticulum (ER) membrane. For this purpose, the Nef sequence was fused to the C-terminal domain of mammalian ER cytochrome b5, a long-lived, tail-anchored (TA) protein. This consistently increased Nef accumulation by more than threefold in many independent transgenic tobacco plants. Real-time polymerase chain reaction of mRNA levels and protein pulse-chase analysis indicated that the increase was not caused by higher transcript levels but by enhanced protein stability. Subcellular fractionation and immunocytochemistry indicated that Nef-TA accumulated on the ER membrane. Over-expression of mammalian or plant ER cytochrome b5 caused the formation of stacked membrane structures, as observed previously in similar experiments performed in mammalian cells; however, Nef-TA did not alter membrane organization in tobacco cells. Finally, Nef could be removed in vitro by its tail-anchor, taking advantage of an engineered thrombin cleavage site. These results open up the way to use tail-anchors to improve foreign protein stability in the plant cytosol without perturbing cellular functions. [source]

Evaluation of a vertical frozen soil barrier at oak ridge national laboratory

REMEDIATION, Issue 3 2000
Stanley W. Lynn
Arctic Foundations, Inc. (AFI), of Anchorage, Alaska, has developed a freeze barrier system designed to hydraulically isolate a contaminant source area. The system can be used for long-term or temporary containment of groundwater until appropriate remediation techniques can be applied. The technology was evaluated under the United States Environmental Protection Agency's (EPA's) Superfund Innovative Technology Evaluation (SITE) program at the United States Department of Energy's (DOE's) Oak Ridge National Laboratory (ORNL) facility in Oak Ridge, Tennessee. For the demonstration, an array of freeze pipes called "thermoprobes" was installed to a depth of 30 feet below ground surface around a former waste collection pond and keyed into bedrock. The system was used to establish an impermeable frozen soil barrier to hydraulically isolate the pond. Demonstration personnel collected independent data to evaluate the technology's performance. A variety of evaluation tools were used,including a groundwater dye tracing investigation, groundwater elevation measurements, and subsurface soil temperature data,to determine the effectiveness of the freeze barrier system in preventing horizontal groundwater flow beyond the limits of the frozen soil barrier. Data collected during the demonstration provided evidence that the frozen soil barrier was effective in hydraulically isolating the pond. [source]

FREEZE: A Celebration of Design in the Modern North

Eric Firley
Abstract Brian Carter reports from Anchorage, in Alaska, where he co-curated an art and architectural event in early 2009 that set out to celebrate the climate, landscape and light of the icy north. Copyright © 2009 John Wiley & Sons, Ltd. [source]

Combined role of type IX collagen and cartilage oligomeric matrix protein in cartilage matrix assembly: Cartilage oligomeric matrix protein counteracts type IX collagen,induced limitation of cartilage collagen fibril growth in mouse chondrocyte cultures

K. Blumbach
Objective Defects in the assembly and composition of cartilage extracellular matrix are likely to result in impaired matrix integrity and increased susceptibility to cartilage degeneration. The aim of this study was to determine the functional interaction of the collagen fibril,associated proteins type IX collagen and cartilage oligomeric matrix protein (COMP) during cartilage matrix formation. Methods Primary chondrocytes from mice deficient in type IX collagen and COMP (double-deficient) were cultured in monolayer or alginate beads. Anchorage of matrix proteins, proteoglycan and collagen content, collagen crosslinks, matrix metalloproteinase activity, and mechanical properties of the matrix were measured. Electron microscopy was used to study the formation of fibrillar structures. Results In cartilage lacking both type IX collagen and COMP, matrilin 3 showed decreased matrix anchorage. Less matrilin 3 was deposited in the matrix of double-deficient chondrocytes, while larger amounts were secreted into the medium. Proteoglycans were less well retained in the matrix formed in alginate cultures, while collagen deposition was not significantly affected. Electron microscopy revealed similar cartilage collagen fibril diameters in the cultures of double-deficient and wild-type chondrocytes. In contrast, a larger fibril diameter was observed in the matrix of chondrocytes deficient in only type IX collagen. Conclusion Our results show that type IX collagen and COMP are involved in matrix assembly by mediating the anchorage and regulating the distribution of other matrix macromolecules such as proteoglycans and matrilins and have counteracting effects on collagen fibril growth. Loss of type IX collagen and COMP leads to matrix aberrations that may make cartilage more susceptible to degeneration. [source]

Anchorage of Titanium Implants with Different Surface Characteristics: An Experimental Study in Rabbits

Klaus Gotfredsen DDS
ABSTRACT Purpose: To compare the anchorage of titanium implants with different surface roughness and topography and to examine histologically the peri-implant bone after implant removal. Materials and Methods: Screw implants with five different surface topographies were examined: (1) turned ("machined"), (2) TiO2 -blasted with particles of grain size 10 to 53 ,m; (3) TiO2 -blasted, grain size 63 to 90 ,m; (4) TiO2 -blasted, grain size 90 to 125 ,m; (5) titanium plasma-sprayed (TPS). The surface topography was determined by the use of an optical instrument. Twelve rabbits, divided into two groups, had a total of 120 implants inserted in the tibiae. One implant from each of the five surface categories was placed within the left tibia of each rabbit. By a second operation, implants were installed in the right tibia, after 2 weeks in group A and after 3 weeks in group B. Fluorochrome labeling was performed after 1 and 3 weeks. Removal torque (RMT) tests of the implants were performed 4 weeks after the second surgery in group A and 9 weeks after the second surgery in group B. Thus, in group A, two healing groups were created, representing 4 and 6 weeks, respectively. The corresponding healing groups in group B were 9 and 12 weeks. The tibiae were removed, and each implant site was dissected, fixed, and embedded in light-curing resin. Ground sections were made, and the peri-implant bone was analyzed using fluorescence and light microscopy. Results: The turned implants had the lowest Sa and Sy values, whereas the highest scores were recorded for the TPS implants. The corresponding Sa and Sy values for the TiO2 -blasted implants were higher when a larger size of grain particles had been used for blasting. At all four observation intervals, the TPS implants had the highest and the turned implants the lowest RMT scores. The differences between the various TiO2 -blasted implants were, in general, small, but the screws with the largest Sa value had higher RMT scores at 6, 9, and 12 weeks than implants with lower Sa values. The histologic analysis of the sections representing 6, 9, and 12 weeks revealed that fractures or ruptures were present in the marginal, cortical peri-implant bone. In such sections representing the TPS and TiO2 -blasted implant categories, ruptures were frequently found in the zone between the old bone and the newly formed bone, as well as within the newly formed bone. Conclusions: The present study demonstrated that a clear relation exists between surface roughness, described in Sa values, and implant anchorage assessed by RMT measurements. The anchorage appeared to increase with the maturation of bone tissue during healing. [source]

Myosin Va phosphorylated on Ser1650 is found in nuclear speckles and redistributes to nucleoli upon inhibition of transcription

CYTOSKELETON, Issue 6 2008
Maria Cristina S. Pranchevicius
Abstract Nuclear actin and nuclear myosins have been implicated in the regulation of gene expression in vertebrate cells. Myosin V is a class of actin-based motor proteins involved in cytoplasmic vesicle transport and anchorage, spindle-pole alignment and mRNA translocation. In this study, myosin-Va, phosphorylated on a conserved serine in the tail domain (phospho-ser1650 MVa), was localized to subnuclear compartments. A monoclonal antibody, 9E6, raised against a peptide corresponding to phosphoserine1650 and flanking regions of the murine myosin Va sequence, was immunoreactive to myosin Va heavy chain in cellular and nuclear extracts of HeLa cells, PC12 cells and B16-F10 melanocytes. Immunofluorescence microscopy with this antibody revealed discrete irregular spots within the nucleoplasm that colocalized with SC35, a splicing factor that earmarks nuclear speckles. Phospho-ser1650 MVa was not detected in other nuclear compartments, such as condensed chromatin, Cajal bodies, gems and perinucleolar caps. Although nucleoli also were not labeled by 9E6 under normal conditions, inhibition of transcription in HeLa cells by actinomycin D caused the redistribution of phospho-ser1650 MVa to nucleoli, as well as separating a fraction of phospho-ser1650 MVa from SC35 into near-neighboring particles. These observations indicate a novel role for myosin Va in nuclear compartmentalization and offer a new lead towards the understanding of actomyosin-based gene regulation. Cell Motil. Cytoskeleton 2008. © 2008 Wiley-Liss, Inc. [source]

Palladin is a novel binding partner for Ena/VASP family members

CYTOSKELETON, Issue 1 2004
Malika Boukhelifa
Abstract Palladin is an actin-associated protein that contains proline-rich motifs within its amino-terminal sequence that are similar to motifs found in zyxin, vinculin, and the Listeria protein ActA. These motifs are known to be potential binding sites for the Vasodilator-Stimulated Phosphoprotein (VASP). Here, we demonstrate that palladin is an additional direct binding partner for VASP, by using co-immunoprecipitation and blot overlay techniques with both endogenous palladin and recombinant myc-tagged palladin. These results show that VASP binds to full-length palladin and also to the amino-terminal half of palladin, where the polyproline motifs are located. Using a synthetic peptide array, two discrete binding sites for VASP were identified within palladin's proline-rich amino-terminal domain. Using double-label immunofluorescence staining of fully-spread and actively-spreading fibroblasts, the extent of co-localization of palladin and VASP was explored. These proteins were found to strongly co-localize along stress fibers, and partially co-localize in focal adhesions, lamellipodia, and focal complexes. These results suggest that the recently described actin-associated protein palladin may play an important role in recruiting VASP to sites of actin filament growth, anchorage, and crosslinking. Cell Motil. Cytoskeleton 58:17,29, 2004. © 2004 Wiley-Liss, Inc. [source]

Hemidesmosome protein dynamics in live epithelial cells

CYTOSKELETON, Issue 2 2003
Daisuke Tsuruta
Abstract Hemidesmosomes mediate stable anchorage of epithelial cells to laminin-5 in the basement membrane zone and have been likened to spot-welds. Indeed, it has been assumed that hemidesmosomes are not dynamic, at least when compared to other matrix adhesion sites including focal contacts. We tested this notion by monitoring the fate of green fluorescent protein (GFP)-tagged human integrin ,4 subunit (GFP-h,4) and GFP-tagged 180-kD human bullous pemphigoid (BP) autoantigen (GFP-BP180) in live cultures of 804G cells that assemble numerous mature hemidesmosomes. In subconfluent 804G cells, both GFP-h,4 and GFP-BP180 protein clusters are not stable but assemble into and disassemble out of cat paw,like arrays at a relatively rapid rate. In confluent populations of 804G cells, although some cat paw,like clusters of both GFP-h,4 and GFP-BP180 are stable over periods of >60 min, other GFP-h,4 and GFP-BP180 protein arrays form and/or disappear during the same time period. Moreover, individual labeled particles show considerable motility in the plane of the membrane. Fluorescence recovery after photobleaching analyses provide a further indication of the dynamics of hemidesmosome proteins. In particular, bleached GFP-h,4 protein clusters in confluent cells recover signal within about 30 min, indicating that there is a relatively rapid turnover of hemidesmosome components in protein arrays clustered along the substratum attached surface of a cell. The rate of recovery is dependent on an intact microfilament system. In sharp contrast, bleached GFP-BP180 protein clusters in confluent cells fail to recover signal even when observed for longer than 60 min. To evaluate hemidesmosome protein dynamics in motile cells, we monitored GFP-h,4 and GFP-BP180 in 804G cells populating scrape wound sites in vitro. In these migratory cells, which lack mature hemidesmosomes, integrin ,4 subunit and BP180 protein clusters progressively assemble and disassemble into linear and cat-paw arrays. In summary, hemidesmosome protein clusters, like their counterparts in focal contacts, are dynamic. We discuss these results in relation to hemidesmosome functions. Cell Motil. Cytoskeleton 54:122,134, 2003. © 2003 Wiley-Liss, Inc. [source]

Crown fragment reattachment: report of an extensive case with intra-canal anchorage

Gustavo M. S. Oliveira
Initially, the fractured crown was splinted to the adjacent teeth with orthodontic wire and composite resin. Subsequently, the crown fragment was reattached by means of a fiber post using a hybrid composite resin. Early stage success was achieved with the observance of normality in function, esthetics, and health of the tooth and surrounding periodontal structures. An athletic mouthguard was fabricated to prevent further trauma. Advantages, disadvantages, and prognosis of the treatment presented are discussed. [source]

Bioreactor for cultivation of red beet hairy roots and in situ recovery of primary and secondary metabolites

Bhagyalakshmi Neelwarne
Abstract To arrive at an appropriate bioreactor design and in situ recovery of the products, red beet hairy roots were used as a model system where the levels of betalain pigments (betacyanins and betaxanthins) were followed as secondary metabolite and the peroxidase enzyme as primary metabolite. Medium volume and other kinetic parameters were found to play significant roles by way of directly affecting the biomass yield rather than a specific metabolite. The hydrodynamic stress created on the roots by large culture volume could be minimized by pulse-feeding of medium in shake-flasks; and by separating the biomass chamber from the aerated medium reservoir in circulatory fed-batch bioreactor. Accordingly the bioreactor was modified to provide anchorage and air-enrichment chamber which resulted in higher formation of both the metabolites than in shake-flasks. Various down-stream processing aspects such as in situ release of pigments by non-destructive methods, followed by adsorption through a column and recovery by desorption were optimized for betalains. A strategy for simultaneous recovery of pigment and peroxidase was worked out using aqueous two phase extraction (ATPE). [source]

Abstention, alcohol use and risk of myocardial infarction in men and women taking account of social support and working conditions: the SHEEP case,control study

ADDICTION, Issue 10 2003
Anders Romelsjö
ABSTRACT Aims, Very few studies indicating that low,moderate alcohol consumption protects from myocardial infarction (MI) controlled for social support and working conditions, which could confound the findings. Therefore, a first aim was to study the risk of non-fatal and total MI in relation to volume of alcohol consumption and measures of social support and working conditions. A second aim was to analyse the impact of the volume of earlier alcohol use in abstainers. Design, Data came from a case,control study, the Stockholm Heart Epidemiology Program (SHEEP), including first MI among Swedish citizens 45,70 years old. Setting, Stockholm County 1992,94. Participants, There were 1095 cases of MI in men and 471 in women (928 and 372 were non-fatal), and 2339 living controls from the general population. Measurement, Information about alcohol use at different periods in life and job strain, social anchorage and life control besides pre-existing health problems, smoking, physical activity, socio-economic status and marital status was obtained by a questionnaire from the cases and the controls. Findings, In multivariate logistic regression analyses, the relative risk for MI (especially non-fatal) was reduced among alcohol consumers. RR for non-fatal MI was 0.52 (95% confidence intervals 0.32, 0.85) in men with a consumption of 50,69.9 g 100% ethanol/day and 0.21 (95% confidence interval 0.06, 0.77) in women with a consumption of 30 g or more per day (reference category 0.1,5 g 100% ethanol/day). Men who were abstainers during the previous 1,10 years and with an earlier average consumption of 5,30 g 100% ethanol/day had a significantly lower relative risk compared to such abstainers with an earlier higher consumption. Earlier consumption among abstainers may also have an impact on gender differences in MI. Analyses showed positive interaction between abstention and low life-control in women, but only 4% of the female cases were due to this interaction. There were no other interactions between measures of alcohol use and social anchorage, life control and working situations. Conclusion, Alcohol use had a protective impact on MI, with little impact of job strain, social anchorage and life control, giving increased support for a protective impact of low-moderate alcohol use. The level of previous alcohol consumption among male 1,10-year-long abstainers influenced the risk of MI. [source]

Selective down-regulation of the ,6-integrin subunit in melanocytes by UVB light

Sven Krengel
Abstract:,In vivo, melanocytes bind to laminin (LM) molecules of the basement membrane (BM) via the integrins ,3,1 and ,6,1, and they adhere to neighbouring keratinocytes via E-cadherin. Only few studies have addressed the impact of ultraviolet (UV) light on the interaction of melanocytes with their microenvironment. In this report, we examined the influence of UVB irradiation on the expression of the most important melanocyte-adhesion molecules (E-, N-cadherin, ,2-, ,3-, ,5-, ,6-, ,V-, ,1-, ,3-integrins and ICAM-1) in vitro by flow cytometry. We were able to demonstrate that the ,6-integrin subunit is selectively and reversibly down-regulated by UVB in a dwzm 150ose-dependent manner. In comparison, keratinocytes lacked UVB-inducible alterations in the expression of ,6-integrin. In the presence of LM-1, the UVB-induced down-regulation of ,6-integrin in melanocytes was significantly reduced. Moreover, LM-1 increased the resistance of melanocytes to UVB-induced cell death, as measured by annexinV-binding analysis. This effect was reversed by preincubation with an ,6-integrin-blocking antibody. By immunofluorescence, we could demonstrate that UVB leads to a dose-dependent internalization of ,6-integrin, providing an obvious explanation for the down-regulation on the outer cell surface observed by flow cytometry. We suggest that adhesion to LM-1 through ,6-integrin represents a protective mechanism for melanocytes to withstand UVB damage. Through ,6-integrin internalization, sunburns might alter the interaction between melanocytes and the BM, resulting in apoptosis induced by loss of anchorage (anoikis). Repeated sunburns may then lead to the selection of a population of melanocytes which are capable of anchorage-independent survival, culminating in solar nevogenesis and melanoma development. [source]

Identity Politics and the Domestic Context of Turkey's European Union Accession

Necati Polat
This article observes a transformation in the largely essentializing, decontextualized form of identity politics that long defined political cosmology in Turkey, now in the process of negotiating accession to the European Union (EU). Accordingly, identity politics , not only the bread and butter of both Kurdish nationalism and Islamism, but also a justification for exhortations towards a limited, authoritarian democracy by Kemalists, the major power holders , is receding in favour of a civic, non-divisive political culture enabled by the EU anchorage. In danger of losing the longstanding centre,periphery configuration in an enhanced, participatory democracy and, concomitant with it, the periphery clientelism created by the waning identity politics, Kemalist nationalists, Islamists and Kurdish separatists appear to have stopped squabbling among themselves and joined forces against Turkey's EU bid. [source]

Evidence for Indo-Roman Trade from Bet Dwarka Waters, West Coast of India

A. S. Gaur
India had a very active maritime trade contact with the Roman world between the 4th century BC and the 4th century AD. In this context recent finds of stone anchors, potsherds, lead anchors and a lead ingot from 5 to 8 m water-depth near Bet Dwarka jetty is significant. The sherds include amphoras, jars, bowls and lids. Archaeological finds along the Indian coast and comparison between amphoras from Bet Dwarka and the Mediterranean suggest that the artefacts from Bet Dwarka may be datable to between the 1st century BC and the 2nd century AD. The numbers of stone anchors suggests that this was an ancient anchorage. © 2005 The Nautical Archaeology Society [source]

Disc structure function and its potential for repair

J. Melrose
The intervertebral disc (IVD) is the largest predominantly avascular, aneural, alymphatic structure of the human body. It provides articulation between adjoining vertebral bodies and also acts as a weight-bearing cushion dissipating axially applied spinal loads. The IVD is composed of an outer collagen-rich annulus fibrosus (AF) and a central proteoglycan (PG)-rich nucleus pulposus (NP). Superior and inferior cartilaginous endplates (CEPs), thin layers of hyaline-like cartilage, cover the ends of the vertebral bodies. The AF is composed of concentric layers (lamellae) which contain variable proportions of type I and II collagen, this tissue has high tensile strength. The NP in contrast is a gelatinous PG-rich tissue which provides weight-bearing properties to the composite disc structure. With the onset of age, cells in the NP progressively die as this tissue becomes depleted of PGs, less hydrated and more fibrous as the disc undergoes an age-dependent fibrocartilaginous transformation. Such age-dependent cellular and matrix changes can decrease the discs' biomechanical competence and trauma can further lead to failure of structural components of the disc. Annular defects are fairly common and include vertebral rim-lesions, concentric (circumferential) annular tears (separation of adjacent annular lamellae) and radial annular tears (clefts which initiate within the NP). While vascular in-growth around annular tears has been noted, evidence from human post-mortem studies indicate they have a limited ability to undergo repair. Several experimental approaches are currently under evaluation for their ability to promote the repair of such annular lesions. These include growth of AF fibrochondrocytes on a resorbable polycaprolactone (PCL) bio-membrane.1 Sheets of fibrochondrocytes lay down type-I collagen and actin stress fibres on PCL. These matrix components are important for the spatial assembly of the collagenous lamella during annular development and correct phenotypic expression of cells in biomatrices.1 An alternative approach employs preparation of tissue engineered IVDs where AF and NP cells are separately cultured in polyglycolic acid and sodium alginate biomatrices, either separately or within a manifold designed to reproduce the required IVD dimensions for its use as a prospective implant device.2 AF and NP cells have also been grown on tissue culture inserts after their recovery from alginate bead culture to form plugs of tissue engineered cartilage.3 A key component in this latter strategy was the stimulation of the high density disc cell cultures with osteogenic protein-1 (OP-1) 200 ng/mL.3 This resulted in the production of tissue engineered AF and NP plugs with compositions, histochemical characteristics and biomechanical properties approaching those of the native disc tissues.2,3 Such materials hold reat promise in future applications as disc or annular implants. The introduction of appropriate genes into disc cells by gene transduction methodology using adenoviral vectors or ,gene-gun' delivery systems also holds considerable promise for the promotion of disc repair processes.4 Such an approach with the OP-1 gene is particularly appealing.5 The anchoring of discal implants to vertebral bodies has also been evaluated by several approaches. A 3D fabric based polyethylene biocomposite holds much promise as one such anchorage device6 while biological glues used to seal fibrocartilaginous structures such as the AF and meniscus8 following surgical intervention, also hold promise in this area. Several very promising new experimental approaches and strategies are therefore currently under evaluation for the improvement of discal repair. The aforementioned IVD defects are a common cause of disc failure and sites of increased nerve in-growth in symptomatic IVDs in man and are thus often sources of sciatic-type pain. Annular defects such as those described above have formerly been considered incapable of undergoing spontaneous repair thus a clear need exists for interventions which might improve on their repair. Based on the rapid rate of progress and the examples outlined above one may optimistically suggest that a successful remedy to this troublesome clinical entity will be developed in the not so distant future. References 1JohnsonWEBet al. (2001) Directed cytoskeletal orientation and intervertebral disc cell growth: towards the development of annular repair techniques. Trans Orthop Res Soc26, 894. 2MizunoHet al. (2001) Tissue engineering of a composite intervertebral disc. Trans Orthop Res Soc26, 78. 3MatsumotoTet al. (2001) Formation of transplantable disc shaped tissues by nucleus pulposus and annulus fibrosus cells: biochemical and biomechanical properties. Trans Orthop Res Soc26, 897. 4NishidaKet al. (2000) Potential applications of gene therapy to the treatment of intervertebral disc disorders. Clin Orthop Rel Res379 (Suppl), S234,S241. 5MatsumotoTet al. (2001) Transfer of osteogenic protein-1 gene by gene gun system promotes matrix synthesis in bovine intervertebral disc and articular cartilage cells. Trans Orthop Res Soc26, 30. 6ShikinamiY , Kawarada (1998) Potential application of a triaxial three-dimensional fabric (3-DF) as an implant. Biomaterials19, 617,35. [source]

From anchorage dependent proliferation to survival: Lessons from redox signalling

IUBMB LIFE, Issue 5 2008
Paola Chiarugi
Abstract Anchorage to extracellular matrix (ECM) is essential for the execution of the mitotic program of nontransformed cells as they need simultaneous signals starting from mitogenic molecules, as growth factors (GFs), and adhesive agents belonging to ECM. Reactive oxygen species play a key function during both GF and integrin receptor signalling and are therefore recognised to have a synergistic function with several others transducers for anchorage-dependent growth (ADG). Indeed, redox-regulated proteins include protein tyrosine phosphatases, protein tyrosine kinases, small GTPases, cytoskeleton proteins, as well as several transcription factors. In this review, we focus on the role of reactive oxygen species (ROS) as key second messengers granting a proper executed mitosis for anchorage-dependent cells through redox regulation of several downstream targets. Besides, redox signals elicited by ECM contact assure a protection from anoikis, a specific apoptosis induced by lack of anchorage. Cancer cells frequently show a deregulation of ROS production and a constitutive oxidative stress has been associated to the achievement of an invasive phenotype. Hence, in cancer cells, the constitutive deregulation of both mitogenic and survival pathways, likely mimicking autocrine/adhesive signals, helps to guide the transformed cells to escape the innate apoptotic response to abolish the signals started by cell/ECM contact, thus sustaining the spreading of anchorage-independent cancer cells and the metastases growth. © 2008 IUBMB IUBMB Life, 60(5): 301,307, 2008 [source]

A study on the sorption of NO3, and F, on the carboxymethylated starch-based hydrogels loaded with Fe2+ ions

Ghanshyam S. Chauhan
Abstract Using the principle of geochemistry of fluoride, green and cost effective anion adsorbents were developed for the removal of F, from water systems. The scheme was further applied for the removal of NO3, also. Carboxymethylated starch functionalized through network formation with acrylamide was used as adsorbent, and the resultant hydrogels were loaded with Fe2+ ions to generate anchorage for the anions. Sorption of Fe2+ was studied as a function of different factors such as time, temperature, pH, and ion strength. The network having the highest Fe2+ uptake was loaded with the Fe2+ ions under optimum conditions and used for the sorption of F, and NO3,. High efficiency has been observed for F,, as even up to 100% uptake has been observed within just 10 minutes. The support shows high selectivity for NO3,, which was used as anion reference. Thermodynamics of sorption confirms low order and low energy processes. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci, 2007 [source]

Characterization of cartilagenous tissue formed on calcium polyphosphate substrates in vitro

Stephen D. Waldman
Abstract Successful joint resurfacing by tissue-engineered cartilage has been limited, in part, by an inability to secure the implant to bone. To overcome this, we have developed the methodology to form a cartilage implant in vitro consisting of a layer of cartilagenous tissue overlying a porous, biodegradable calcium polyphosphate (CPP) substrate. As bone will grow into the CPP after implantation, it will result in anchorage of the cartilage. In this study, the cartilagenous tissue formed in vitro after 8 weeks in culture was characterized and compared to native articular cartilage. Light microscopic examination of histological sections showed that there was a continuous layer of cartilagenous tissue on, and integrated with the subsurface of, the CPP substrate. The in vitro -formed tissue achieved a similar thickness to native articular cartilage (mean ± SEM: in vitro = 0.94 ± 0.03 mm; ex vivo = 1.03 ± 0.01 mm). The cells in the in vitro -formed tissue synthesized large proteoglycans (Kav ± SEM: in vitro = 0.27 ± 0.01; ex vivo = 0.27 ± 0.01) and type II collagen similar to the chondrocytes in the ex-vivo cartilage. The in vitro -formed tissue had a similar amount of proteoglycan (GAG ,g/mg dry wt.: in vitro = 198 ± 10; ex vivo = 201 ± 13) but less collagen than the native cartilage (hydroxyproline ,g/mg dry wt.: in vitro = 21 ± 1; ex vivo = 70 ± 8). The in vitro -formed tissue had only about 3% of the load-bearing capacity and stiffness of the native articular cartilage, determined from unconfined mechanical compression testing. Although low, this was within the range of properties reported by others for tissue-engineered cartilage. It is possible that the limited load-bearing capacity is the result of the low collagen content and further studies are required to identify the conditions that will increase collagen synthesis. © 2002 Wiley Periodicals, Inc. J Biomed Mater Res 62:323,330, 2002 [source]

Resistance to experimental tumorigenesis in cells of a long-lived mammal, the naked mole-rat (Heterocephalus glaber)

AGING CELL, Issue 4 2010
Sitai Liang
Summary The naked mole-rat (NMR, Heterocephalus glaber) is a long-lived mammal in which spontaneous cancer has not been observed. To investigate possible mechanisms for cancer resistance in this species, we studied the properties of skin fibroblasts from the NMR following transduction with oncogenes that cause cells of other mammalian species to form malignant tumors. Naked mole-rat fibroblasts were transduced with a retrovirus encoding SV40 large T antigen and oncogenic RasG12V. Following transplantation of transduced cells into immunodeficient mice, cells rapidly entered crisis, as evidenced by the presence of anaphase bridges, giant cells with enlarged nuclei, multinucleated cells, and cells with large number of chromosomes or abnormal chromatin material. In contrast, similarly transduced mouse and rat fibroblasts formed tumors that grew rapidly without crisis. Crisis was also observed after > 40 population doublings in SV40 TAg/Ras-expressing NMR cells in culture. Crisis in culture was prevented by additional infection of the cells with a retrovirus encoding hTERT (telomerase reverse transcriptase). SV40 TAg/Ras/hTERT-expressing NMR cells formed tumors that grew rapidly in immunodeficient mice without evidence of crisis. Crisis could also be induced in SV40 TAg/Ras-expressing NMR cells by loss of anchorage, but after hTERT transduction, cells were able to proliferate normally following loss of anchorage. Thus, rapid crisis is a response of oncogene-expressing NMR cells to growth in an in vivo environment, which requires anchorage independence, and hTERT permits cells to avoid crisis and to achieve malignant tumor growth. The unique reaction of NMR cells to oncogene expression may form part of the cancer resistance of this species. [source]

The microanatomy of the distal arrector pili: possible role for ,1,1 and ,5,1 integrins in mediating cell-cell adhesion and anchorage to the extracellular matrix

Jeri Kersten Mendelson
The arrector pili (AP) muscle is a small band of smooth muscle that attaches proximally to the bulge area of the pilosebaceous apparatus in the reticular dermis and extends up toward the epidermis. The distal anatomy of the AP and the anchorage mechanism allowing hair erection have not been previously described. Integrins are likely candidates mediating this attachment. Immunohistochemical techniques were used to determine the distribution of the following integrins: ,1, ,2, ,3, ,4, ,5, ,6 and ,1 as well as fibronectin. Frozen human scalp tissue was sectioned in traditional planes, obliquely and horizontally to visualize microanatomy in three dimensions. Histological examination revealed that the distal portions of smooth muscle fibers splay extensively between collagen bundles of the upper dermis. Integrin subunits ,1, ,5 and ,1 were expressed by the AP muscle. Analysis of the relative density of immunoreactivity in digitized sections revealed increased ,5 subunit expression at the extracellular matrix (ECM)-muscle interface. These data suggest that anchorage of the AP muscle to the ECM is via ,5,1 integrin and ,1,1 integrin functions in muscle cell-cell adhesion. Extensive splaying of smooth muscle fibers may allow increased surface area contact between the ECM and smooth muscle cells expressing peripherally situated ,5 integrin. [source]

Implant-Supported Obturator Overdenture for Extensive Maxillary Resection Patient: A Clinical Report

Cláudio Rodrigues Leles DDS
Abstract This clinical report presents an implant-retained obturator overdenture solution for a Prosthodontic Diagnostic Index Class IV maxillectomy patient with a large oronasal communication and severe facial asymmetry, loss of upper lip and midfacial support, severe impairment of mastication, deglutition, phonetics, and speech intelligibility. Due to insufficient bone support to provide satisfactory zygomaticus implant anchorage, conventional implants were placed in the body of the left zygomatic arch and in the right maxillary tuberosity. Using a modified impression technique, a cobalt-chromium alloy framework with three overdenture attachments was constructed to retain a complete maxillary obturator. Patient-reported functional and quality of life measure outcomes were dramatically improved after treatment and at the two-year follow-up. [source]

Corrosion of Dental Magnet Attachments for Removable Prostheses on Teeth and Implants

Arne F. Boeckler DMD, Dr Med Dent
Abstract Purpose: For a long time, the use of magnets for the anchorage of dental prostheses failed due to lack of biocompatibility and the magnets' high susceptibility to corrosion in the mouth. These facts make encapsulation of the magnetic alloy with a corrosion-resistant, tight, and functionally firm sealing necessary. Due to different products and analysis methods, it is not feasible to compare the findings for contemporary products with the sparse and rather old test results in the literature. Therefore, the aim of this study was the standardized control and the comparison of the corrosion behavior of modern magnetic attachments for use on teeth and dental implants. Materials and Methods: Thirty-seven components of magnetic attachments on implants and natural teeth from different alloys (NdFeB, SmCo, Ti, CrMoMnTiFe, etc.) as delivered by the manufacturers or fabricated according to their instructions were examined for their corrosion behavior using the statical immersion analysis (ISO 10271:2001). Four specimens of every product with the same design were used. An uncased SmCo magnet served as control. Analyses after 1, 4, 7, and 28 days of the storage in corrosion solution were made. The eluate was examined quantitatively on the alloy components of the respective component with the help of optical emission spectrometry (,g/cm2). The results were compared to the requirements of ISO standard 22674:2006. In addition, existing corrosion products were also defined in the solution after 28 days. The results were analyzed descriptively and statistically to determine possible significant differences (t -test and Mann-Whitney-Wilcoxon rank-sums test; p < 0.05). Results: Dissolved metal ions could be found on all tested products. The release after 1 and 4 days was different for all specimens. In the group of implant abutments, the highest ion release after 7 days was found (all measurements ,g/cm2): Fe (13.94, Magfit-IP-IDN dome type), Pd (1.53, Medical-anchor), Cr (1.32, Magfit-IP-IDN dome type), Ti (1.09, Magfit-IP-IDN abutment), Co (0.81, Medical-anchor), and B (0.6, Magfit-IP-IDN dome type). After 28 days, the analyzed ion release increased irregularly: Fe (173.58, Magfit-IP-IDN dome type), Pd (44.17, Medical-anchor), Cr (2.02, Magfit-IP-IDN dome type), Ti (2.11, Magfit-IP-IDN abutment), Co (26.13, Medical-anchor), B (1.77, Magfit-IP-IDN dome type), and Nd (79.18, Magfit-IP-IDN dome type). In the group of magnetic systems on natural teeth, the highest ion release after 7 days was found for Fe (4.81, Magfit DX 800 keeper), Cr (1.18, Magfit DX 800 keeper), Pd (0.21, Direct System Keeper), Ni (0.18, WR-Magnet S3 small), Co (0.12, Direct System Keeper), and Ti (0.09, Magna Cap , Mini). After 28 days, the analyzed ion release increased non-uniformly: Fe (31.92, Magfit DX 800 Keeper), Cr (6.65, Magfit DX 800 Keeper), Pd (18.19, Direct System Keeper), Ni (0.61, WR-Magnet S3 small), Co (10.94, Direct System Keeper), Ti (0.83, Magna Cap , Mini), and Pd (2.78, EFM Alloy). In contrast, the uncased control magnet showed an exponential release after 7 days of Sm ions (55.06) and Co-ions (86.83), after 28 days of Sm ions (603.91) and Co ions (950.56). The release of corrosion products of all tested products stayed significantly under the limit of 200 ,g/cm2 (ISO 22674:2006). In contrast, the non-encapsulated control magnet exceeded that limit significantly. Conclusion: The analysis of the corrosion behavior of modern magnetic attachments for use on teeth and dental implants according to ISO 10271:2001 showed that metal ions had dissolved on all specimens. In the case of one product, the magnet corroded. For this product, an improvement of the capsulation would be desirable. None of the products reached the limit specified in ISO 22674:2006. All products seem to be suitable for dental application. Further studies in regard to the specific biocompatibility and possible cytotoxic effects on mucosa and tissue would be desirable. [source]

Continuous supply of TGF,3 via adenoviral vector promotes type I collagen and viability of fibroblasts in alginate hydrogel

Yongchang Yao
Abstract In recent years, transforming growth factor-,3 (TGF,3) has interested more and more researchers with its competence in engineered histogenesis. In the present study we employed recombinant adenoviral vectors to deliver the constitutively active TGF,3 gene to human dermal fibroblasts, which could maintain the continuous secretion of TGF,3 from the cells. The expression of type I collagen in the Ad-TGF,3 group increased significantly in comparison with other three groups: Neg (cells without treatment of the adenovirus), Ad-null (cells with treatment of the adenovirus, without the inserted gene) and Ad-shRNA (cells with treatment of the adenovirus encoding shRNA specific for type I collagen). Additionally, we demonstrated that TGF,3 enhanced the expression of Smad4 while inhibiting that of MMP-9, thus promoting the collagen transcription via the Smad signal transduction pathway and restraining collagen degradation by MMP-9, which contributed to the increasing type I collagen expression level. As type I collagen mediates cell,material interactions by providing anchorage, the viability of encapsulated fibroblasts in Ad-TGF,3 group was significantly higher than that in other three groups. Accordingly, this approach forms an effective way to improve the compatibility of non-adhesive hydrogels containing anchorage-dependent cells. Copyright © 2010 John Wiley & Sons, Ltd. [source]

Cell transformation induced by hepatitis C virus NS3 serine protease

R. Zemel
Persistent infection with hepatitis C virus (HCV) may lead to hepatocellular carcinoma (HCC). It has been suggested that HCV-encoded proteins are directly involved in the tumorigenic process. The HCV nonstructural protein NS3 has been identified as a virus-encoded serine protease. To study whether HCV NS3 has oncogenic activity, nontumorigenic rat fibroblast (RF) cells were stably transfected with an expression vector containing cDNA for the NS3 serine protease (nucleotides 3356,4080). The NS3 serine protease activity was determined in the transfected cells. The transfected cells grew rapidly and proliferated serum independently, lost contact inhibition, grew anchorage independently in soft agar and induced significant tumour formation in nude mice. Cells transfected with an expression vector containing a mutated NS3 serine protease (serine 139 to alanine at the catalytic site) showed no transforming abilities; their growth was dependent on serum and they did not grow anchorage independently in soft agar. Moreover, cells transfected with the NS3 serine protease and treated with the chymotrypsin inhibitors TPCK and PMSF (a serine protease inhibitor) lost their transforming feature. These results suggest that the NS3 serine protease of HCV is involved in cell transformation and that the ability to transform requires an active enzyme. [source]

The interdental gingiva, a visible guide for placement of mini-implants

YS Chun
Structured Abstract Authors,,, Chun YS, Lee SK, Wikesjö UME, Lim WH Objectives,,, To determine whether the tip of the interdental gingiva can serve as a visible guide for placement of mini-implants. Setting and Sample population,,, Computer tomography (CT) images from 15 males and 15 females (mean age 27 years, range: 23,35 years) were used to evaluate the distance from the tip of the interdental gingiva to the alveolar crest from the central incisor to the 1st molar. The distance from a reference point to the tip of interdental gingiva was recorded from study models using a caliper. The distance between the reference point and the alveolar crest was recorded using CT and added to the model recordings thus providing the distance from the tip of interdental gingiva to the alveolar crest for the various interdental sites. Two-way anova and Student,Newman,Keuls test for multiple comparisons were used for the statistical analysis. Results,,, There was no significant difference in the distance from the tip of interdental gingiva to the alveolar crest between maxilla and mandible. The distance between the tip of interdental gingiva and the alveolar crest at the central/lateral incisors was the shortest compared with that of other sites. There was also a statistically significant difference between the male and female groups except for the maxillary 2nd premolar/1st molar interradicular site. Conclusion,,, The tip of interdental gingiva appears a reasonable visual guide for the placement of mini-implants for orthodontic anchorage. [source]

AKT2 is a downstream target of metabotropic glutamate receptor 1 (Grm1)

Seung-Shick Shin
Summary We reported earlier on the oncogenic properties of Grm1 by demonstrating that stable Grm1 -mouse-melanocytic clones proliferate in the absence of growth supplement and anchorage in vitro. In addition, these clones also exhibit aggressive tumorigenic phenotypes in vivo with short latency in tumor formation in both immunodeficient and syngeneic mice. We also detected strong activation of AKT in allograft tumors specifically AKT2 as the predominant isoform involved. In parallel, we assessed several human melanoma biopsy samples and found again that AKT2 was the predominantly activated AKT in these human melanoma biopsies. In cultured stable Grm1 -mouse-melanocytic clones, as well as an metabotropic glutamate receptor 1 (Grm1) expressing human melanoma cell line, C8161, stimulation of Grm1 by its agonist led to the activation of AKT, while preincubation with Grm1-antagonist abolished Grm1-agonist-induced AKT activation. In addition, a reduction in tumor volume of Grm1 -mouse-melanocytic-allografts was detected in the presence of small interfering AKT2 RNA (siAKT2). Taken together, these results showed that, in addition to the MAPK pathway previously reported being a downstream target of stimulated Grm1, AKT2 is another downstream target in Grm1 mediated melanocyte transformation. [source]

Structural features for the mechanism of antitumor action of a dimeric human pancreatic ribonuclease variant

Antonello Merlino
Abstract A specialized class of RNases shows a high cytotoxicity toward tumor cell lines, which is critically dependent on their ability to reach the cytosol and to evade the action of the ribonuclease inhibitor (RI). The cytotoxicity and antitumor activity of bovine seminal ribonuclease (BSRNase), which exists in the native state as an equilibrium mixture of a swapped and an unswapped dimer, are peculiar properties of the swapped form. A dimeric variant (HHP2-RNase) of human pancreatic RNase, in which the enzyme has been engineered to reproduce the sequence of BSRNase helix-II (Gln28,Leu, Arg31,Cys, Arg32,Cys, and Asn34,Lys) and to eliminate a negative charge on the surface (Glu111,Gly), is also extremely cytotoxic. Surprisingly, this activity is associated also to the unswapped form of the protein. The crystal structure reveals that on this molecule the hinge regions, which are highly disordered in the unswapped form of BSRNase, adopt a very well-defined conformation in both subunits. The results suggest that the two hinge peptides and the two Leu28 side chains may provide an anchorage to a transient noncovalent dimer, which maintains Cys31 and Cys32 of the two subunits in proximity, thus stabilizing a quaternary structure, similar to that found for the noncovalent swapped dimer of BSRNase, that allows the molecule to escape RI and/or to enhance the formation of the interchain disulfides. [source]