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Sample records for chondrocyte grafts four-year

  1. The INNOVATION Trial: four-year safety and effectiveness of the INCRAFT® AAA Stent-Graft System for endovascular repair.

    Science.gov (United States)

    Pratesi, Giovanni; Pratesi, Carlo; Chiesa, Roberto; Coppi, Gioacchino; Scheinert, Dierk; Brunkwall, Jan S; van der Meulen, Stefaan; Torsello, Giovanni

    2017-10-01

    This paper reports the 4-year safety and effectiveness of the INCRAFT® AAA Stent-Graft System (Cordis Corp., Milpitas, CA, USA), an ultra-low-profile device for the treatment of abdominal aortic aneurysms. The INNOVATION Trial is the prospective, first-in-human, multicenter trial to evaluate the safety and effectiveness of the INCRAFT® System. Patients underwent annual clinical and computed tomography angiography examination as part of the study protocol. The INCRAFT® AAA Stent-Graft System is a customizable tri-modular design, with an ultra-low profile (14-Fr) delivery system. Patient were treated under approved protocol, the prescribed clinical and imaging follow-up at annually through 5 years. Results analyzed and adjudicated by a clinical events committee, independent core laboratory, and a data safety and monitoring board. This manuscript reports results through 4 years of follow-up. A total of 60 patients were enrolled in the trial, all of whom were successfully treated. Follow-up rates at 1 and 4 years were 93% (56/60) and 85% (51/60), respectively. All-cause mortality at 4 years was 17.6% and no death was AAA-, device-, or procedure-related. The secondary reintervention rate at 1 year was 4.6%, primarily the result of stent thrombosis. In total, 10 patients required 13 post-procedure interventions within 4-years of follow-up (2 to repair a type I endoleak, 4 to repair a type II endoleak, 1 for stent thrombosis, 1 for renal stenosis, 1 for aneurysm enlargement, 2 for limb migration and 2 for prosthesis stenosis or occlusion). There were 4 cases (10%) of aneurysm enlargement reported at the 4 year follow-up. At 4 years, 38 out of 39 patients were free from type I and III endoleaks. There were no proximal type I or type III endoleaks at 4-year follow-up. Core laboratory evaluation of the postoperative imaging studies indicated absence of endograft migration while a single fracture was demonstrated without any clinical sequelae. The INCRAFT® AAA Stent-Graft

  2. Engineering cartilaginous grafts using chondrocyte-laden hydrogels supported by a superficial layer of stem cells.

    Science.gov (United States)

    Mesallati, Tariq; Buckley, Conor T; Kelly, Daniel J

    2017-05-01

    During postnatal joint development, progenitor cells that reside in the superficial region of articular cartilage first drive the rapid growth of the tissue and later help direct the formation of mature hyaline cartilage. These developmental processes may provide directions for the optimal structuring of co-cultured chondrocytes (CCs) and multipotent stromal/stem cells (MSCs) required for engineering cartilaginous tissues. The objective of this study was to engineer cartilage grafts by recapitulating aspects of joint development where a population of superficial progenitor cells drives the development of the tissue. To this end, MSCs were either self-assembled on top of CC-laden agarose gels (structured co-culture) or were mixed with CCs before being embedded in an agarose hydrogel (mixed co-culture). Porcine infrapatellar fat pad-derived stem cells (FPSCs) and bone marrow-derived MSCs (BMSCs) were used as sources of progenitor cells. The DNA, sGAG and collagen content of a mixed co-culture of FPSCs and CCs was found to be lower than the combined content of two control hydrogels seeded with CCs and FPSCs only. In contrast, a mixed co-culture of BMSCs and CCs led to increased proliferation and sGAG and collagen accumulation. Of note was the finding that a structured co-culture, at the appropriate cell density, led to greater sGAG accumulation than a mixed co-culture for both MSC sources. In conclusion, assembling MSCs onto CC-laden hydrogels dramatically enhances the development of the engineered tissue, with the superficial layer of progenitor cells driving CC proliferation and cartilage ECM production, mimicking certain aspects of developing cartilage. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

  3. Stimulation of chondrocyte proliferation following photothermal, thermal, and mechanical injury in ex-vivo cartilage grafts

    Science.gov (United States)

    Pandoh, Nidhi S.; Truong, Mai T.; Diaz-Valdes, Sergio H.; Gardiner, David M.; Wong, Brian J.

    2002-06-01

    Laser irradiation may stimulate chondrocytes proliferation in the peripheral region surrounding a photothermally-heated area in rabbit nasal septal cartilage. In this study, ex- vivo rabbit nasal septal cartilages maintained in culture were irradiated with an Nd:YAG laser ((lambda) equals1.32 micrometers , 4-16 sec, 10-45 W/cm2) to examine the relationship between the diameter of replicating cells and irradiation time. Also, this study investigated whether proliferation occurs following heating (by immersion in hot saline baths, with a heated metal rod, and a soldering iron) and mechanical modification (crushing with a metal stamp and scoring with a scalpel). Replicating chondrocytes were identified using a Bromodeoxyuridine (BrdU) double antibody detection system in whole mount tissue. Light microscopy was used to confirm the presence of BrdU stained chondrocytes. The mechanical and thermal stressors used failed to produce a proliferative response in chondrocytes as previously seen with laser irradiation. We suspect that chondrocyte proliferation may be induced as a response to alteration in matrix structure produced by photothermal, thermal, or mechanical modification of the matrix. Heat generated by a laser to stimulate chondrocyte proliferation may lead to new treatment options for degenerative articular diseases and disorders. Laser technology can be adapted for use with minimally invasive surgical instrumentation to deliver light into otherwise inaccessible regions of the body.

  4. Four-Year Itch

    Science.gov (United States)

    Pierce, Dennis

    2017-01-01

    Ohio's decision to let community colleges award four-year degrees is part of a growing national trend. When this article went to press, more than 90 community colleges across 19 states offered active four-year degree programs. Counting New York's Fashion Institute of Technology, which technically is a community college, but offers degrees as high…

  5. A comparison of self-assembly and hydrogel encapsulation as a means to engineer functional cartilaginous grafts using culture expanded chondrocytes.

    Science.gov (United States)

    Mesallati, Tariq; Buckley, Conor T; Kelly, Daniel J

    2014-01-01

    Despite an increased interest in the use of hydrogel encapsulation and cellular self-assembly (often termed "self-aggregating" or "scaffold-free" approaches) for tissue-engineering applications, to the best of our knowledge, no study to date has been undertaken to directly compare both approaches for generating functional cartilaginous grafts. The objective of this study was to directly compare self-assembly (SA) and agarose hydrogel encapsulation (AE) as a means to engineer such grafts using passaged chondrocytes. Agarose hydrogels (5 mm diameter × 1.5 mm thick) were seeded with chondrocytes at two cell seeding densities (900,000 cells or 4 million cells in total per hydrogel), while SA constructs were generated by adding the same number of cells to custom-made molds. Constructs were either supplemented with transforming growth factor (TGF)-β3 for 6 weeks, or only supplemented with TGF-β3 for the first 2 weeks of the 6 week culture period. The SA method was only capable of generating geometrically uniform cartilaginous tissues at high seeding densities (4 million cells). At these high seeding densities, we observed that total sulphated glycosaminoglycan (sGAG) and collagen synthesis was greater with AE than SA, with higher sGAG retention also observed in AE constructs. When normalized to wet weight, however, SA constructs exhibited significantly higher levels of collagen accumulation compared with agarose hydrogels. Furthermore, it was possible to engineer such functionality into these tissues in a shorter timeframe using the SA approach compared with AE. Therefore, while large numbers of chondrocytes are required to engineer cartilaginous grafts using the SA approach, it would appear to lead to the faster generation of a more hyaline-like tissue, with a tissue architecture and a ratio of collagen to sGAG content more closely resembling native articular cartilage.

  6. Regulative mechanisms of chondrocyte adhesion

    DEFF Research Database (Denmark)

    Schmal, Hagen; Mehlhorn, Alexander T; Fehrenbach, Miriam

    2006-01-01

    Interaction between chondrocytes and extracellular matrix is considered a key factor in the generation of grafts for matrix-associated chondrocyte transplantation. Therefore, our objective was to study the influence of differentiation status on cellular attachment. Adhesion of chondrocytes...... to collagen type II increased after removal from native cartilage up to the third day in monolayer in a dose-dependent manner. Following dedifferentiation after the second passage, adhesion to collagen types I (-84%) and II (-46%) decreased, whereas adhesion to fibrinogen (+59%) and fibronectin (+43......%) increased. A cartilage construct was developed based on a clinically established collagen type I scaffold. In this matrix, more than 80% of the cells could be immobilized by mechanisms of adhesion, filtration, and cell entrapment. Confocal laser microscopy revealed focal adhesion sites as points of cell...

  7. Grafting

    Energy Technology Data Exchange (ETDEWEB)

    Garnett, J L [New South Wales Univ., Kensington (Australia). School of Chemistry

    1979-01-01

    The unique value of ionizing radiation for the initiation of grafting to backbone polymers is discussed. The principles of the technique are briefly reviewed. The conditions under which free radicals and ions participate in these reactions are examined. Examples of representative grafting processes are considered to illustrate where the technique can be of potential commercial value to a wide range of industries. The general principles of these grafting reactions are shown to be applicable to radiation induced rapid cure technology such as is provided by electron beam processing facilities. Grafting reactions initiated by UV are also treated and shown to be of importance because of the many similarities in properties of the ionizing radiation and UV systems, also the rapid industrial exploitation of EB and sensitized UV processing technology. Possible future trends in radiation grafting are outlined.

  8. Four years after the JCO criticality accident

    International Nuclear Information System (INIS)

    Sumita, Kenji

    2003-01-01

    It has been about four years since the first criticality accident in Japan. The JCO accident site was not so far from this auditorium. I have been asked to give a short review of important results from the various technical investigations on the accident that have been performed during the past four years. I will also give a short introduction to the changes that have been made in the nuclear safety regulation systems of the Japanese Government. (author)

  9. Importance of Donor Chondrocyte Viability for Osteochondral Allografts.

    Science.gov (United States)

    Cook, James L; Stannard, James P; Stoker, Aaron M; Bozynski, Chantelle C; Kuroki, Keiichi; Cook, Cristi R; Pfeiffer, Ferris M

    2016-05-01

    Osteochondral allograft (OCA) transplantation provides a biological treatment option for functional restoration of large articular cartilage defects in multiple joints. While successful outcomes after OCA transplantation have been linked to viable donor chondrocytes, the importance of donor cell viability has not been comprehensively validated. To use a canine model to determine the importance of donor chondrocyte viability at the time of implantation with respect to functional success of femoral condylar OCAs based on radiographic, gross, cell viability, histologic, biochemical, and biomechanical outcome measures. Controlled laboratory study. After approval was obtained from the institutional animal care and use committee, adult female dogs (N = 16) were implanted with 8-mm cylindrical OCAs from male dogs in the lateral and medial femoral condyles of 1 knee. OCAs were preserved for 28 or 60 days after procurement, and chondrocyte viability was quantified before implantation. Two different storage media, temperatures, and time points were used to obtain a spectrum of percentage chondrocyte viability at the time of implantation. A successful outcome was defined as an OCA that was associated with graft integration, maintenance of hyaline cartilage, lack of associated cartilage disorder, and lack of fibrillation, fissuring, or fibrous tissue infiltration of the allograft based on subjective radiographic, gross, and histologic assessments at 6 months after implantation. Chondrocyte viability ranged from 23% to 99% at the time of implantation. All successful grafts had >70% chondrocyte viability at the time of implantation, and no graft with chondrocyte viability <70% was associated with a successful outcome. Live-dead stained sections and histologic findings with respect to cell morphological features suggested that successful grafts were consistently composed of viable chondrocytes in lacunae, while grafts that were not successful were composed of nonviable

  10. Elastic cartilage reconstruction by transplantation of cultured hyaline cartilage-derived chondrocytes.

    Science.gov (United States)

    Mizuno, M; Takebe, T; Kobayashi, S; Kimura, S; Masutani, M; Lee, S; Jo, Y H; Lee, J I; Taniguchi, H

    2014-05-01

    Current surgical intervention of craniofacial defects caused by injuries or abnormalities uses reconstructive materials, such as autologous cartilage grafts. Transplantation of autologous tissues, however, places a significant invasiveness on patients, and many efforts have been made for establishing an alternative graft. Recently, we and others have shown the potential use of reconstructed elastic cartilage from ear-derived chondrocytes or progenitors with the unique elastic properties. Here, we examined the differentiation potential of canine joint cartilage-derived chondrocytes into elastic cartilage for expanding the cell sources, such as hyaline cartilage. Articular chondrocytes are isolated from canine joint, cultivated, and compared regarding characteristic differences with auricular chondrocytes, including proliferation rates, gene expression, extracellular matrix production, and cartilage reconstruction capability after transplantation. Canine articular chondrocytes proliferated less robustly than auricular chondrocytes, but there was no significant difference in the amount of sulfated glycosaminoglycan produced from redifferentiated chondrocytes. Furthermore, in vitro expanded and redifferentiated articular chondrocytes have been shown to reconstruct elastic cartilage on transplantation that has histologic characteristics distinct from hyaline cartilage. Taken together, cultured hyaline cartilage-derived chondrocytes are a possible cell source for elastic cartilage reconstruction. Crown Copyright © 2014. Published by Elsevier Inc. All rights reserved.

  11. Four years in / four years out : two university buildings 2000-2008

    Energy Technology Data Exchange (ETDEWEB)

    Lovell, J. [Royal Inst. of British Architects, London (United Kingdom); Washington Univ., St. Louis, MO (United States). Graduate School of Architecture and Urban Design

    2009-07-01

    The University of Cambridge commissioned London-based Allies and Morrison to develop and design 2 new buildings for its Sidgwick campus, England. Two naturally ventilated buildings were completed and occupied in 2004. The Estate Management and Building Service at the University of Cambridge is now reviewing the actual energy performance of both buildings in terms of user comfort. The finding will be compared against original projections of the design team. The review will also compare actual running costs against projected costs and feedback from building users and facilities management. The approach taken by client and design team towards the life-time energy performance of the building was examined in terms of comfort and control and load management. A more complex system of sub-metering is now proposed for all new buildings. Post occupancy feedback data was used to evaluate the successes and failures of the project teams in terms of energy performance, thermal comfort and building management communication four years after completion. This paper noted that if viewed over 30 years, the initial construction budget of a building accounts for only a fraction of the true cost of a building. During its lifetime, primary expenditure will likely to be upon personnel costs; work performance and salaries, staff retention and absenteeism. Therefore, the success of a building must be seen in the context of its whole life and the comfort of its users. 14 refs., 9 figs.

  12. Access to Four-Year Public Colleges and Degree Completion

    OpenAIRE

    Joshua Goodman; Michael Hurwitz; Jonathan Smith

    2015-01-01

    Does access to four-year colleges affect degree completion for students who would otherwise attend two-year colleges? Admission to Georgia’s four-year public sector requires minimum SAT scores. Regression discontinuity estimates show that access to this sector increases four-year college enrollment and college quality, largely by diverting students from two-year colleges. Access substantially increases bachelor’s degree completion rates for these relatively low-skilled students. SAT retaking ...

  13. Scaffold-assisted cartilage tissue engineering using infant chondrocytes from human hip cartilage.

    Science.gov (United States)

    Kreuz, P C; Gentili, C; Samans, B; Martinelli, D; Krüger, J P; Mittelmeier, W; Endres, M; Cancedda, R; Kaps, C

    2013-12-01

    Studies about cartilage repair in the hip and infant chondrocytes are rare. The aim of our study was to evaluate the use of infant articular hip chondrocytes for tissue engineering of scaffold-assisted cartilage grafts. Hip cartilage was obtained from five human donors (age 1-10 years). Expanded chondrocytes were cultured in polyglycolic acid (PGA)-fibrin scaffolds. De- and re-differentiation of chondrocytes were assessed by histological staining and gene expression analysis of typical chondrocytic marker genes. In vivo, cartilage matrix formation was assessed by histology after subcutaneous transplantation of chondrocyte-seeded PGA-fibrin scaffolds in immunocompromised mice. The donor tissue was heterogenous showing differentiated articular cartilage and non-differentiated tissue and considerable expression of type I and II collagens. Gene expression analysis showed repression of typical chondrocyte and/or mesenchymal marker genes during cell expansion, while markers were re-induced when expanded cells were cultured in PGA-fibrin scaffolds. Cartilage formation after subcutaneous transplantation of chondrocyte loaded PGA-fibrin scaffolds in nude mice was variable, with grafts showing resorption and host cell infiltration or formation of hyaline cartilage rich in type II collagen. Addition of human platelet rich plasma (PRP) to cartilage grafts resulted robustly in formation of hyaline-like cartilage that showed type II collagen and regions with type X collagen. These results suggest that culture of expanded and/or de-differentiated infant hip cartilage cells in PGA-fibrin scaffolds initiates chondrocyte re-differentiation. The heterogenous donor tissue containing immature chondrocytes bears the risk of cartilage repair failure in vivo, which may be possibly overcome by the addition of PRP. Copyright © 2013 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

  14. Repair of experimentally produced defects in rabbit articular cartilage by autologous chondrocyte transplantation

    International Nuclear Information System (INIS)

    Grande, D.A.; Pitman, M.I.; Peterson, L.; Menche, D.; Klein, M.

    1989-01-01

    Using the knee joints of New Zealand White rabbits, a baseline study was made to determine the intrinsic capability of cartilage for healing defects that do not fracture the subchondral plate. A second experiment examined the effect of autologous chondrocytes grown in vitro on the healing rate of these defects. To determine whether any of the reconstituted cartilage resulted from the chondrocyte graft, a third experiment was conducted involving grafts with chondrocytes that had been labeled prior to grafting with a nuclear tracer. Results were evaluated using both qualitative and quantitative light microscopy. Macroscopic results from grafted specimens displayed a marked decrease in synovitis and other degenerative changes. In defects that had received transplants, a significant amount of cartilage was reconstituted (82%) compared to ungrafted controls (18%). Autoradiography on reconstituted cartilage showed that there were labeled cells incorporated into the repair matrix

  15. 2009-2012 State-ANDRA four-year contract

    International Nuclear Information System (INIS)

    2009-01-01

    Signed by State representatives and representatives of the French national Agency for radioactive waste management (ANDRA), this document defines for the four years to come several objectives for the action and the activities of the ANDRA considered as an industrial operator, as an innovating actor possessing a high level research capacity, as a public expert guaranteeing the radioactive waste management solutions, as an information and valorization centre for France and foreign countries. All these objectives are further detailed in appendix

  16. Nuclear Electric - four years on and right on course

    International Nuclear Information System (INIS)

    Hawley, R.

    1994-01-01

    Since the Government's decision, four years ago, to withdraw the nuclear stations from the electricity supply industry privatisation, and place a moratorium on the construction of further nuclear plant, the newly formed nuclear generators, Nuclear Electric and Scottish Nuclear, have risen to the challenge of proving that nuclear power can be as competitive in the United Kingdom as it is elsewhere in the world. The Chief Executive of Nuclear Electric documents changes which have taken place within the organisation since privatisation, presenting data on output and market share and productivity to back his argument. (UK)

  17. Video-EEG recording: a four-year clinical audit.

    LENUS (Irish Health Repository)

    O'Rourke, K

    2012-02-03

    In the setting of a regional neurological unit without an epilepsy surgery service as in our case, video-EEG telemetry is undertaken for three main reasons; to investigate whether frequent paroxysmal events represent seizures when there is clinical doubt, to attempt anatomical localization of partial seizures when standard EEG is unhelpful, and to attempt to confirm that seizures are non-epileptic when this is suspected. A clinical audit of all telemetry performed over a four-year period was carried out, in order to determine the clinical utility of this aspect of the service and to determine means of improving effectiveness in the unit. Analysis of the data showed a high rate of negative studies with no attacks recorded. Of the positive studies approximately 50% showed non-epileptic attacks. Strategies for improving the rate of positive investigations are discussed.

  18. Analyzing the Bitcoin Network: The First Four Years

    Directory of Open Access Journals (Sweden)

    Matthias Lischke

    2016-03-01

    Full Text Available In this explorative study, we examine the economy and transaction network of the decentralized digital currency Bitcoin during the first four years of its existence. The objective is to develop insights into the evolution of the Bitcoin economy during this period. For this, we establish and analyze a novel integrated dataset that enriches data from the Bitcoin blockchain with off-network data such as business categories and geo-locations. Our analyses reveal the major Bitcoin businesses and markets. Our results also give insights on the business distribution by countries and how businesses evolve over time. We also show that there is a gambling network that features many very small transactions. Furthermore, regional differences in the adoption and business distribution could be found. In the network analysis, the small world phenomenon is investigated and confirmed for several subgraphs of the Bitcoin network.

  19. Suicide in India: a four year retrospective study.

    Science.gov (United States)

    Mohanty, Sachidananda; Sahu, Geeta; Mohanty, Manoj Kumar; Patnaik, Manju

    2007-05-01

    Suicide is one of the ten leading causes of death in the world, accounting for more than a million deaths annually. The purpose of the study was to identify the risk groups. In the four-year period from January 2000 to December 2003, 588 suicide victims were autopsied. Information was obtained by interviewing the acquaintances of the victim, perusal of hospital records and the autopsy findings. All the cases were analyzed as to sex, age, and methods of suicide, seasonal variation, diurnal variation and other sociological aspects. The present study depicts a suicidal rate of 11.76 per 100,000 population. Males and females were almost equally the sufferers. The largest number of victims were found in the age group of 21-30 years. Hanging and poisoning constituted the two major modes of suicides (63%). Majority of the victims were mentally sound, married and were from rural background. Victims were mostly drawn from low socioeconomic status (48%). Less educated or illiterates were usually the victims. Suicidal note was detected in 5% of cases. Suicidal tendency and alcohol intake could not be encountered in most of the cases. Indoor incidence was almost double of the outdoor incidence, mostly seen in rainy season (43%) and occurred almost equally during day and night. Financial burden (37%) and marital disharmony (35%) were the principal reasons for the suicide.

  20. FACT. More than four years of blazar monitoring

    Energy Technology Data Exchange (ETDEWEB)

    Dorner, Daniela [Universitaet Wuerzburg (Germany); Collaboration: FACT-Collaboration

    2016-07-01

    Since October 2011, the First G-APD Cherenkov Telescope (FACT) has been collecting more than 5500 hours of physics data. Thanks to the silicon based photosensors (SiPMs, aka G-APDs), observations during bright ambient light like full moon can be carried out without degradation of the sensors. Keeping the gain of the SiPMs stable using an online feedback system, a stable and homogeneous detector performance is achieved. Based on this and an automatic data taking procedure, an unbiased longterm data sample is collected. An automatic quick look analysis provides results shortly after the data are taken allowing to send flare alerts within the same night. The main targets for FACT are the bright TeV blazars Mrk 421 and Mrk 501 which are monitored since January 2012. In addition, several other sources like for example the Crab Nebula, 1ES 1959+650, 1ES 2344+54.1 are observed. In this presentation, the results from more than four years of monitoring are summarized. Several flares from Mrk 501 and Mrk 421 are discussed in the multi-wavelength (MWL) context. Mrk 501 underwent major outbursts in June 2012 and June 2014 during the yearly MWL campaigns. Mrk 421 showed a bright flare in April 2013 where also MWL observations are available. 1ES 1959+650 showed enhanced flux in autumn 2015. Results from these observations are discussed.

  1. Postural changes in dental hygienists. Four-year longitudinal study.

    Science.gov (United States)

    Barry, R M; Woodall, W R; Mahan, J M

    1992-01-01

    Numerous surveys identify the occurrence of musculoskeletal complaints as a concern in dentistry. However, no longitudinal data exist to indicate whether postural changes occur as a result of practicing dental hygiene. The purpose of this preliminary, four-year longitudinal study was to investigate whether any postural changes developed during the hygienists' clinical education and/or during subsequent dental hygiene practice after one and/or two years. It was anticipated that the awkward positions and intense physical demands placed on hygienists might initiate musculoskeletal problems, but that no postural changes would occur over this short period of time. Nine of 10 dental hygienists in the graduating class of 1987 were surveyed for existing musculoskeletal complaints, and the subjects were photographed for a measurement of postural change. Responses from participants indicated an increase in musculoskeletal-related complaints in each of the six areas investigated. The photographic findings indicated that one of the nine hygienists showed an increase in forward head posture, a postural change.

  2. Cognitive Effects of Technology Over Four Years of College

    Directory of Open Access Journals (Sweden)

    Chad N. Loes

    2016-05-01

    Full Text Available Technology permeates higher education, yet less is known about the use of established technologies, such as email and other electronic communication mediums (e.g., discussion boards, listservs for instructional purposes on important student outcomes such as cognitive development. In this study, we use data from the Wabash National Study to estimate the effects of email and other electronic medium use for academic purposes on three measures of cognitive development over four years of college. To investigate this, we regress each measure of cognitive development on email and electronic medium use, while simultaneously controlling for a wide array of potential confounding influences. Net of these influences, we find that email and electronic medium use are positively associated with gains in students’ Need for Cognition. These same technologies fail to have more than a chance influence on students’ critical thinking skills, however. Lastly, email use is associated with gains in the Positive Attitudes Toward Literacy measure for Whites and females, whereas electronic medium use leads to gains in the same outcome for racial and ethnic minorities. While institutions consider newer technologies for instructional purposes, our findings suggest established technologies can play a powerful role in the development of students’ cognitive skills.

  3. Four Years on Orbit at the Moon with LOLA

    Science.gov (United States)

    Smith, D. E.; Zuber, M. T.; Neumann, G. A.; Mazarico, E.; Torrence, M. H.; Lemoine, F. G.

    2013-12-01

    After four years of near-continuous operation at the Moon, the Lunar Orbiter Laser Altimeter (LOLA) continues to collect altimetry, surface roughness, slope and normal reflectance data. Although the instrument is beginning to show the effects of tens of thousands of thermal cycles and the natural process of the aging of the laser transmitters, LOLA continues to acquire data on the sunlit portion of every orbit on all 5 laser beams when below 100-km altitude. LOLA has acquired over 6x10^9 altimeter measurements, all geodetically controlled to the center-of-mass of the Moon with a radial precision of around 10 cm and an accuracy of about 1 meter. The position of the measurements on the lunar surface is primarily limited by the knowledge of the position of the spacecraft in orbit; in the last year the LRO orbit accuracy has improved significantly as a result of the availability of an accurate gravity model of the Moon from the GRAIL Discovery mission. Our present estimate of positional accuracy is less than 10 m rms but is only achievable with a GRAIL gravity model to at least degree and order 600 because of the perturbing gravitational effect of the Moon's surface features. Significant improvements in the global shape and topography have assisted the Lunar Reconnaissance Orbiter Camera (LROC) stereo mapping program, and the identification of potential lunar landing sites for ESA and Russia, particularly in the high-latitude polar regions where 5- and 10-meter average horizontal resolution has been obtained. LOLA's detailed mapping of the polar regions has improved the delineation of permanently-shadowed areas and assisted in the understanding of the LEND neutron data and its relationship to surface slopes. Recently, a global, calibrated LOLA normal albedo dataset at 1064 nm has been developed and is being combined with analysis and modeling by the Diviner team for the identification of the coldest locations in the polar regions.

  4. Fate of Meckel's cartilage chondrocytes in ocular culture

    International Nuclear Information System (INIS)

    Richman, J.M.; Diewert, V.M.

    1988-01-01

    Modulation of the chondrocyte phenotype was observed in an organ culture system using Meckel's cartilage. First branchial arch cartilage was dissected from fetal rats of 16- and 17-day gestation. Perichondrium was mechanically removed, cartilage was split at the rostral process, and each half was grafted into the anterior chamber of an adult rat eye. The observed pattern of development in nonirradiated specimens was the following: hypertrophy of the rostral process and endochondral-type ossification, fibrous atrophy in the midsection, and mineralization of the malleus and incus. A change in matrix composition of the implanted cartilage was demonstrated with immunofluorescence staining for cartilage-specific proteoglycan (CSPG). After 15 days of culture, CSPG was found in the auricular process but not in the midsection or rostral process. In order to mark the implanted cells and follow their fate, cartilage was labeled in vitro with [3H]thymidine [3H]TdR). Immediately after labeling 20% of the chondrocytes contained [3H]TdR. After culturing for 5 days, 20% of the chondrocytes were still labeled and 10% of the osteogenic cells also contained radioactive label. The labeling index decreased in both cell types with increased duration of culture. Multinucleated clast-type cells did not contain label. Additional cartilages not labeled with [3H]TdR were exposed to between 20000 and 6000 rad of gamma irradiation before ocular implantation. Irradiated cartilage did not hypertrophy or form bone but a fibrous region developed in the midsection. Cells of the host animal were not induced to form bone around the irradiated cartilage. Our studies suggest that fully differentiated chondrocytes of Meckel's cartilage have the capacity to become osteocytes, osteoblasts, and fibroblasts

  5. Skin graft

    Science.gov (United States)

    Skin transplant; Skin autografting; FTSG; STSG; Split thickness skin graft; Full thickness skin graft ... donor site. Most people who are having a skin graft have a split-thickness skin graft. This takes ...

  6. Evolution of Autologous Chondrocyte Repair and Comparison to Other Cartilage Repair Techniques

    Directory of Open Access Journals (Sweden)

    Ashvin K. Dewan

    2014-01-01

    Full Text Available Articular cartilage defects have been addressed using microfracture, abrasion chondroplasty, or osteochondral grafting, but these strategies do not generate tissue that adequately recapitulates native cartilage. During the past 25 years, promising new strategies using assorted scaffolds and cell sources to induce chondrocyte expansion have emerged. We reviewed the evolution of autologous chondrocyte implantation and compared it to other cartilage repair techniques. Methods. We searched PubMed from 1949 to 2014 for the keywords “autologous chondrocyte implantation” (ACI and “cartilage repair” in clinical trials, meta-analyses, and review articles. We analyzed these articles, their bibliographies, our experience, and cartilage regeneration textbooks. Results. Microfracture, abrasion chondroplasty, osteochondral grafting, ACI, and autologous matrix-induced chondrogenesis are distinguishable by cell source (including chondrocytes and stem cells and associated scaffolds (natural or synthetic, hydrogels or membranes. ACI seems to be as good as, if not better than, microfracture for repairing large chondral defects in a young patient’s knee as evaluated by multiple clinical indices and the quality of regenerated tissue. Conclusion. Although there is not enough evidence to determine the best repair technique, ACI is the most established cell-based treatment for full-thickness chondral defects in young patients.

  7. Articular chondrocyte metabolism and osteoarthritis

    Energy Technology Data Exchange (ETDEWEB)

    Leipold, H.R.

    1989-01-01

    The three main objectives of this study were: (1) to determine if depletion of proteoglycans from the cartilage matrix that occurs during osteoarthritis causes a measurable increase of cartilage proteoglycan components in the synovial fluid and sera, (2) to observe what effect intracellular cAMP has on the expression of matrix components by chondrocytes, and (3) to determine if freshly isolated chondrocytes contain detectable levels of mRNA for fibronectin. Canine serum keratan sulfate and hyaluronate were measured to determine if there was an elevation of these serum glycosaminoglycans in a canine model of osteoarthritis. A single intra-articular injection of chymopapain into a shoulder joint increased serum keratan sulfate 10 fold and hyaluronate less than 2 fold in 24 hours. Keratan sulfate concentrations in synovial fluids of dogs about one year old were unrelated to the presence of spontaneous cartilage degeneration in the joints. High keratan sulfate in synovial fluids correlated with higher keratan sulfate in serum. The mean keratan sulfate concentration in sera of older dogs with osteoarthritis was 37% higher than disease-free controls, but the difference between the groups was not statistically significant. Treatment of chondrocytes with 0.5 millimolar (mM) dibutyryl cAMP (DBcAMP) caused the cells to adopt a more rounded morphology. There was no difference between the amount of proteins synthesized by cultures treated with DBcAMP and controls. The amount of fibronectin (FN) in the media of DBcAMP treated cultures detected by an ELISA was specifically reduced, and the amount of {sup 35}S-FN purified by gelatin affinity chromatography decreased. Moreover, the percentage of FN containing the extra domain. A sequence was reduced. Concomitant with the decrease in FN there was an increase in the concentration of keratan sulfate.

  8. Cartilage grafting in facial reconstruction with special consideration of irradiated grafts

    International Nuclear Information System (INIS)

    Donald, P.J.

    1986-01-01

    The search for the perfect facial implant for reconstruction of the face continues. Cartilage, once thought to be an undesirable graft material because of its propensity for absorption, has regained popularity in the past decade. Various preparation techniques have been employed to ensure graft sterility and diminished absorption. An improved understanding of cartilage structure and physiology has shed considerable light on the host-graft relationship. Gamma irradiation is a time-honored method of preservation. An experiment was undertaken to investigate the physiology of irradiated cartilage grafts following prolonged implantation on the facial skeleton of sheep and dog. Merthiolate preserved grafts were used as controls. Direct observation, histochemical techniques, autoradiography, and transmission electron micrography were used to determine chondrocyte viability and matrix composition. It was surprising to note that following implantation of 16 to 72 months, complete resorption was seen in 87.7% of the irradiated grafts and in 43.8% of the Merthiolate stored controls. Many of the grafts acquired chondrocytes from the host and produced new proteoglycan matrix as well as undergoing some degree of ossification. A comparison to the clinical situation in humans is made. 98 references

  9. In-vitro chondrogenic potential of synovial stem cells and chondrocytes allocated for autologous chondrocyte implantation

    DEFF Research Database (Denmark)

    Kubosch, Eva Johanna; Heidt, Emanuel; Niemeyer, Philipp

    2017-01-01

    Purpose: The use of passaged chondrocytes is the current standard for autologous chondrocyte implantation (ACI). De-differentiation due to amplification and donor site morbidity are known drawbacks highlighting the need for alternative cell sources. Methods: Via clinically validated flow cytometry...... analysis, we compared the expression of human stem cell and cartilage markers (collagen type 2 (Col2), aggrecan (ACAN), CD44) of chondrocytes (CHDR), passaged chondrocytes for ACI (CellGenix™), bone marrow derived mesenchymal stem cells (BMSC), and synovial derived stem cells (SDSC). Results: Primary...

  10. ROCK inhibitor prevents the dedifferentiation of human articular chondrocytes

    International Nuclear Information System (INIS)

    Matsumoto, Emi; Furumatsu, Takayuki; Kanazawa, Tomoko; Tamura, Masanori; Ozaki, Toshifumi

    2012-01-01

    Highlights: ► ROCK inhibitor stimulates chondrogenic gene expression of articular chondrocytes. ► ROCK inhibitor prevents the dedifferentiation of monolayer-cultured chondrocytes. ► ROCK inhibitor enhances the redifferentiation of cultured chondrocytes. ► ROCK inhibitor is useful for preparation of un-dedifferentiated chondrocytes. ► ROCK inhibitor may be a useful reagent for chondrocyte-based regeneration therapy. -- Abstract: Chondrocytes lose their chondrocytic phenotypes in vitro. The Rho family GTPase ROCK, involved in organizing the actin cytoskeleton, modulates the differentiation status of chondrocytic cells. However, the optimum method to prepare a large number of un-dedifferentiated chondrocytes is still unclear. In this study, we investigated the effect of ROCK inhibitor (ROCKi) on the chondrogenic property of monolayer-cultured articular chondrocytes. Human articular chondrocytes were subcultured in the presence or absence of ROCKi (Y-27632). The expression of chondrocytic marker genes such as SOX9 and COL2A1 was assessed by quantitative real-time PCR analysis. Cellular morphology and viability were evaluated. Chondrogenic redifferentiation potential was examined by a pellet culture procedure. The expression level of SOX9 and COL2A1 was higher in ROCKi-treated chondrocytes than in untreated cells. Chondrocyte morphology varied from a spreading form to a round shape in a ROCKi-dependent manner. In addition, ROCKi treatment stimulated the proliferation of chondrocytes. The deposition of safranin O-stained proteoglycans and type II collagen was highly detected in chondrogenic pellets derived from ROCKi-pretreated chondrocytes. Our results suggest that ROCKi prevents the dedifferentiation of monolayer-cultured chondrocytes, and may be a useful reagent to maintain chondrocytic phenotypes in vitro for chondrocyte-based regeneration therapy.

  11. ROCK inhibitor prevents the dedifferentiation of human articular chondrocytes

    Energy Technology Data Exchange (ETDEWEB)

    Matsumoto, Emi [Department of Orthopaedic Surgery, Science of Functional Recovery and Reconstruction, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, 2-5-1 Shikatacho, Kitaku, Okayama 700-8558 (Japan); Furumatsu, Takayuki, E-mail: matino@md.okayama-u.ac.jp [Department of Orthopaedic Surgery, Science of Functional Recovery and Reconstruction, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, 2-5-1 Shikatacho, Kitaku, Okayama 700-8558 (Japan); Kanazawa, Tomoko; Tamura, Masanori; Ozaki, Toshifumi [Department of Orthopaedic Surgery, Science of Functional Recovery and Reconstruction, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, 2-5-1 Shikatacho, Kitaku, Okayama 700-8558 (Japan)

    2012-03-30

    Highlights: Black-Right-Pointing-Pointer ROCK inhibitor stimulates chondrogenic gene expression of articular chondrocytes. Black-Right-Pointing-Pointer ROCK inhibitor prevents the dedifferentiation of monolayer-cultured chondrocytes. Black-Right-Pointing-Pointer ROCK inhibitor enhances the redifferentiation of cultured chondrocytes. Black-Right-Pointing-Pointer ROCK inhibitor is useful for preparation of un-dedifferentiated chondrocytes. Black-Right-Pointing-Pointer ROCK inhibitor may be a useful reagent for chondrocyte-based regeneration therapy. -- Abstract: Chondrocytes lose their chondrocytic phenotypes in vitro. The Rho family GTPase ROCK, involved in organizing the actin cytoskeleton, modulates the differentiation status of chondrocytic cells. However, the optimum method to prepare a large number of un-dedifferentiated chondrocytes is still unclear. In this study, we investigated the effect of ROCK inhibitor (ROCKi) on the chondrogenic property of monolayer-cultured articular chondrocytes. Human articular chondrocytes were subcultured in the presence or absence of ROCKi (Y-27632). The expression of chondrocytic marker genes such as SOX9 and COL2A1 was assessed by quantitative real-time PCR analysis. Cellular morphology and viability were evaluated. Chondrogenic redifferentiation potential was examined by a pellet culture procedure. The expression level of SOX9 and COL2A1 was higher in ROCKi-treated chondrocytes than in untreated cells. Chondrocyte morphology varied from a spreading form to a round shape in a ROCKi-dependent manner. In addition, ROCKi treatment stimulated the proliferation of chondrocytes. The deposition of safranin O-stained proteoglycans and type II collagen was highly detected in chondrogenic pellets derived from ROCKi-pretreated chondrocytes. Our results suggest that ROCKi prevents the dedifferentiation of monolayer-cultured chondrocytes, and may be a useful reagent to maintain chondrocytic phenotypes in vitro for chondrocyte

  12. The influence of cell-matrix attachment and matrix development on the micromechanical environment of the chondrocyte in tissue-engineered cartilage

    NARCIS (Netherlands)

    Khoshgoftar, M.; Ito, K.; Donkelaar, C.C. van

    2014-01-01

    Insufficiency of mechanical properties of tissue-engineered (TE) cartilage grafts is still a limiting factor for their clinical application. It has been shown that mechanostimulation of chondrocytes enhances synthesis of extracellular matrix (ECM) and thereby improves the mechanical properties of

  13. Oxygen effects on senescence in chondrocytes and mesenchymal stem cells: consequences for tissue engineering.

    Science.gov (United States)

    Moussavi-Harami, Farid; Duwayri, Yazan; Martin, James A; Moussavi-Harami, Farshid; Buckwalter, Joseph A

    2004-01-01

    Primary isolates of chondrocytes and mesenchymal stem cells are often insufficient for cell-based autologous grafting procedures, necessitating in vitro expansion of cell populations. However, the potential for expansion is limited by cellular senescence, a form of irreversible cell cycle arrest regulated by intrinsic and extrinsic factors. Intrinsic mechanisms common to most somatic cells enforce senescence at the so-called "Hayflick limit" of 60 population doublings. Termed "replicative senescence", this mechanism prevents cellular immortalization and suppresses oncogenesis. Although it is possible to overcome the Hayflick limit by genetically modifying cells, such manipulations are regarded as prohibitively dangerous in the context of tissue engineering. On the other hand, senescence associated with extrinsic factors, often called "stress-induced" senescence, can be avoided simply by modifying culture conditions. Because stress-induced senescence is "premature" in the sense that it can halt growth well before the Hayflick limit is reached, growth potential can be significantly enhanced by minimizing culture related stress. Standard culture techniques were originally developed to optimize the growth of fibroblasts but these conditions are inherently stressful to many other cell types. In particular, the 21% oxygen levels used in standard incubators, though well tolerated by fibroblasts, appear to induce oxidative stress in other cells. We reasoned that chondrocytes and MSCs, which are adapted to relatively low oxygen levels in vivo, might be sensitive to this form of stress. To test this hypothesis we compared the growth of MSC and chondrocyte strains in 21% and 5% oxygen. We found that incubation in 21% oxygen significantly attenuated growth and was associated with increased oxidant production. These findings indicated that sub-optimal standard culture conditions sharply limited the expansion of MSC and chondrocyte populations and suggest that cultures for

  14. MR appearance of autologous chondrocyte implantation in the knee: correlation with the knee features and clinical outcome

    Energy Technology Data Exchange (ETDEWEB)

    Takahashi, Tomoki [Department of Radiology and Institute of Orthopaedics, Oswestry, Shropshire (United Kingdom); Kumamoto University, Department of Orthopaedic and Neuro-Musculoskeletal Surgery, Kumamoto (Japan); Tins, Bernhard; McCall, Iain W.; Ashton, Karen [Department of Radiology and Institute of Orthopaedics, Oswestry, Shropshire (United Kingdom); Robert Jones and Agnes Hunt Orthopaedic Hospital NHS Trust, Department of Diagnostic Imaging, Oswestry, Shropshire (United Kingdom); Richardson, James B. [Department of Radiology and Institute of Orthopaedics, Oswestry, Shropshire (United Kingdom); RJAH Orthopaedic Hospital, Institute of Orthopaedics, Oswestry, Shropshire (United Kingdom); Takagi, Katsumasa [Department of Radiology and Institute of Orthopaedics, Oswestry, Shropshire (United Kingdom); Kumamoto Aging Research Institute, Kumamoto (Japan)

    2006-01-01

    To relate the magnetic resonance imaging (MRI) appearance of autologous chondrocyte implantation (ACI) in the knee in the 1st postoperative year with other knee features on MRI and with clinical outcome. Forty-nine examinations were performed in 49 patients at 1 year after ACI in the knee. Forty-one preoperative magnetic resonance (MR) examinations were also available. The grafts were assessed for smoothness, thickness in comparison with that of adjacent cartilage, signal intensity, integration to underlying bone and adjacent cartilage, and congruity of subchondral bone. Presence of overgrowth and bone marrow appearance beneath the graft were also assessed. Presence of osteophyte formation, further cartilage defects, appearance of the cruciate ligaments and the menisci were also recorded. An overall graft score was constructed, using the graft appearances. This was correlated with the knee features and the Lysholm score, a clinical self-assessment score. The data were analysed by a Kruskal-Wallis H test followed by a Mann-Whitney U test with Bonferroni correction as post-hoc test. Of 49 grafts, 32 (65%) demonstrated complete defect filling 1 year postoperatively. General overgrowth was seen in eight grafts (16%), and partial overgrowth in 13 grafts (26%). Bone marrow change underneath the graft was seen; oedema was seen in 23 grafts (47%), cysts in six grafts (12%) and sclerosis in two grafts (4%). Mean graft score was 8.7 (of maximal 12) (95% CI 8.0-9.5). Knees without osteophyte formation or additional other cartilage defects (other than the graft site) had a significantly higher graft score than knees with multiple osteophytes (P=0.0057) or multiple further cartilage defects (P=0.014). At 1 year follow-up improvement in the clinical scores was not significantly different for any subgroup. (orig.)

  15. A spike at benefit exhaustion: Still possible after four years of unemployment

    DEFF Research Database (Denmark)

    Lyk-Jensen, Stéphanie; Weatherall, Cecilie Dohlmann

    European policymakers wonder whether the long-term unemployed can still find jobs if the benefit period is shortened. We investigate this question by analysing how long-term unemployed react to the threat of running out of unemployment insurance (UI) after receiving it for almost four years....... The empirical analysis is based on very precise administrative records of men’s unemployment spells. To identify the effect of UI exhaustion, we exploit a legislative change in the duration of benefits that progressively reduced UI entitlement from five to four years. Our results show a spike 3-6 months before...

  16. Chondrocyte-seeded type I/III collagen membrane for autologous chondrocyte transplantation

    DEFF Research Database (Denmark)

    Niemeyer, Philipp; Lenz, Philipp; Kreuz, Peter C

    2010-01-01

    PURPOSE: We report the 2-year clinical results and identify prognostic factors in patients treated with autologous chondrocyte transplantation by use of a collagen membrane to seed the chondrocytes (ACT-CS). METHODS: This is a prospective study of 59 patients who were treated with ACT......-CS represents a technical modification of membrane-associated autologous chondrocyte transplantation that combines easy handling and attractive application properties with reliable clinical results 24 months after surgery, especially in patients with isolated cartilage defects. Even though the failure rate...

  17. Autologous Chondrocyte Implantation in Osteoarthritic Surroundings

    DEFF Research Database (Denmark)

    Ossendorff, Robert; Grad, Sibylle; Stoddart, Martin J

    2018-01-01

    BACKGROUND: Autologous chondrocyte implantation (ACI) fails in up to 20% of cases. Advanced intra-articular degeneration paired with an inflammatory environment may be closely related to implantation failure. Certain cytokines have been identified to play a major role during early osteoarthritis....... PURPOSE: To investigate the effects of tumor necrosis factor α (TNFα) and its potential inhibition by adalimumab on cartilage regeneration in an in vitro model of ACI. STUDY DESIGN: Controlled laboratory study. METHODS: Bovine articular chondrocytes were cultivated and transferred at passage 3 to fibrin...

  18. Four years experience with filtration systems in commercial nurseries for eliminating Phytophthora species from recirculation water

    Science.gov (United States)

    T. Ufer; M. Posner; H.-P. Wessels; S. Wagner; K. Kaminski; T. Brand; Werres S.

    2008-01-01

    In a four year project, three different filtration systems were tested under commercial nursery conditions to eliminate Phytophthora spp. from irrigation water. Five nurseries were involved in the project. Slow sand filtration systems were tested in three nurseries. In the fourth nursery, a filtration system with lava grains (Shieer® Bio filtration)...

  19. A Critical Reflection: Foster Care Youth Experiences at a Four Year Postsecondary Institution

    Science.gov (United States)

    Greer, Renada D.

    2016-01-01

    Foster care youth face significant challenges to postsecondary educational success, especially while enrolled at four-year institutions. Foster care youth are absent of family support that their non-foster peers receive throughout the college experience. Without family support, foster care youth encounter greater challenges to persevere through…

  20. Building a four year post-secondary curriculum in educational science and technology

    NARCIS (Netherlands)

    Plomp, T.; Verhagen, Pleunes Willem

    1982-01-01

    This report begins by providing background information on educational technology in the Netherlands, differences between a four-year program in the Netherlands and undergraduate programs in the United States, and the structure of Dutch university programs. The need for a Department of Educational

  1. Four Years of Reports of Alcohol-Related Harm to Pediatricians in the Netherlands

    NARCIS (Netherlands)

    van Hoof, Joris Jasper; van Zanten, Eva; van der Lely, Nicolaas

    2015-01-01

    Over the four years of the study, the number of adolescents treated with alcohol-related harm increased significantly (from 297 in 2007 to 684 in 2010), up to a total of 1,616. The dominant reason for hospitalization was “alcohol intoxication” (in total 1,350; 88% of all cases). The gender ratio did

  2. Four-Year Results of a Youth Smoking Prevention Program Using Assertiveness Training.

    Science.gov (United States)

    Del Greco, Linda; And Others

    1986-01-01

    Seventh graders (N=161) participated in health education classes consisting of either an innovative smoking education program, the program plus assertiveness training, or a traditional smoking education program. Data collected four years later revealed no significant differences in smoking behavior, changes in assertion, or sex differences among…

  3. Enhancing Software Engineering Education through Open Source Projects: Four Years of Students' Perspectives

    NARCIS (Netherlands)

    Papadopoulos, P.M.; Stamelos, I.G.; Meiszner, A.

    2015-01-01

    This paper presents the results after four years of running of an instructional method that utilizes free/libre open source software (FLOSS) projects as tools for teaching software engineering in formal education. In the last four academic years, a total of 408 juniors majoring in Informatics (in a

  4. Development of health over four years among middle-aged and older Europeans

    DEFF Research Database (Denmark)

    Vestergaard, Sonja; Eriksen, Mette Lindholm; Andersen-Ranberg, Karen

    2013-01-01

    Older adults in Eastern and Central European countries have a higher risk of developing poor self-rated health over four years. The same geographical pattern is seen for a higher risk of developing hypertension and diabetes. Low educational level significantly increases the risk of developing poor...... self-perceived health....

  5. Positive Youth Development through an Outdoor Physical Activity Programme: Evidence from a Four-Year Evaluation

    Science.gov (United States)

    Armour, Kathleen; Sandford, Rachel

    2013-01-01

    In 2006, Sandford, Armour and Warmington undertook a comprehensive review of the literature on the role of physical activity/sport and physical education in promoting positive development for disaffected youth. This paper revisits the findings of the literature review in light of data from a four-year evaluation of one corporate-sponsored physical…

  6. Analyzing Four-Year Public University and Two-Year College Graduation Rates

    Science.gov (United States)

    Ober, David R.; Beekman, John A.; Pierce, Rebecca L.

    2018-01-01

    This paper examines the graduation rates between 2000 and 2015 of United States colleges and universities at the national, state, and institutional levels. This research focuses on two-year and four-year programs. Rates are investigated longitudinally along with variables that distinguish between public/private institutions, percentages of…

  7. The Pros and Cons of Two-Year Versus Four-Year Degrees.

    Science.gov (United States)

    Urbaniak, Anthony

    1985-01-01

    Reports results of a research survey conducted to determine what differences (job titles, income, relevance of courses, satisfaction, suitability) exist between two-year (Associate of Science) graduates and four-year (Bachelor of Science) graduates in business. Statistical tables are included. (CT)

  8. The differential prevalence of obesity and related behaviors in two vs. four-year colleges.

    Science.gov (United States)

    Laska, Melissa Nelson; Pasch, Keryn E.; Lust, Katherine; Story, Mary; Ehlinger, Ed

    2012-01-01

    The objective of this study was to determine whether obesity prevalence and weight-related behaviors (e.g., diet, physical activity) differ among students enrolled in two-year community/technical colleges and those attending four-year colleges/universities. This information could inform the development of intervention strategies. Through an existing surveillance system of Minnesota post-secondary education institutions, survey data were collected from 16,539 students from 27 campuses (14 two-year college campuses, 13 four-year college/university campuses; 2007-2008), including self-reported physical activity, media use, dietary patterns, weight control behaviors, height and weight. Unadjusted analyses indicated that students enrolled in two-year colleges, particularly females, had a higher prevalence of overweight/obesity, lower levels of physical activity, more television viewing, higher intakes of soda, fast food, and diet pills compared to students attending four-year colleges (pstudents there were fewer differences between two-year and four-year colleges. Controlling for sociodemographic factors (e.g., race/ethnicity, age), most disparities in prevalence estimates remained, though many were attenuated. Overall, few young adults engage in weight-related behaviors consistent with national recommendations. Two-year college students may represent a particularly at-risk group. Disparities between two- and four-year college students exist beyond the sociodemographic differences in these populations. Effective weight-related interventions are needed for young adults, particularly females attending two-year colleges and all males attending post-secondary institutions. PMID:20966910

  9. Skin Graft

    OpenAIRE

    Shimizu, Ruka; Kishi, Kazuo

    2012-01-01

    Skin graft is one of the most indispensable techniques in plastic surgery and dermatology. Skin grafts are used in a variety of clinical situations, such as traumatic wounds, defects after oncologic resection, burn reconstruction, scar contracture release, congenital skin deficiencies, hair restoration, vitiligo, and nipple-areola reconstruction. Skin grafts are generally avoided in the management of more complex wounds. Conditions with deep spaces and exposed bones normally require the use o...

  10. Cartilage repair: Generations of autologous chondrocyte transplantation

    International Nuclear Information System (INIS)

    Marlovits, Stefan; Zeller, Philip; Singer, Philipp; Resinger, Christoph; Vecsei, Vilmos

    2006-01-01

    Articular cartilage in adults has a limited capacity for self-repair after a substantial injury. Surgical therapeutic efforts to treat cartilage defects have focused on delivering new cells capable of chondrogenesis into the lesions. Autologous chondrocyte transplantation (ACT) is an advanced cell-based orthobiologic technology used for the treatment of chondral defects of the knee that has been in clinical use since 1987 and has been performed on 12,000 patients internationally. With ACT, good to excellent clinical results are seen in isolated post-traumatic lesions of the knee joint in the younger patient, with the formation of hyaline or hyaline-like repair tissue. In the classic ACT technique, chondrocytes are isolated from small slices of cartilage harvested arthroscopically from a minor weight-bearing area of the injured knee. The extracellular matrix is removed by enzymatic digestion, and the cells are then expanded in monolayer culture. Once a sufficient number of cells has been obtained, the chondrocytes are implanted into the cartilage defect, using a periosteal patch over the defect as a method of cell containment. The major complications are periosteal hypertrophy, delamination of the transplant, arthrofibrosis and transplant failure. Further improvements in tissue engineering have contributed to the next generation of ACT techniques, where cells are combined with resorbable biomaterials, as in matrix-associated autologous chondrocyte transplantation (MACT). These biomaterials secure the cells in the defect area and enhance their proliferation and differentiation

  11. Cartilage repair: Generations of autologous chondrocyte transplantation

    Energy Technology Data Exchange (ETDEWEB)

    Marlovits, Stefan [Department of Traumatology, Center for Joint and Cartilage, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna (Austria)]. E-mail: stefan.marlovits@meduniwien.ac.at; Zeller, Philip [Department of Traumatology, Center for Joint and Cartilage, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna (Austria); Singer, Philipp [Department of Traumatology, Center for Joint and Cartilage, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna (Austria); Resinger, Christoph [Department of Traumatology, Center for Joint and Cartilage, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna (Austria); Vecsei, Vilmos [Department of Traumatology, Center for Joint and Cartilage, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna (Austria)

    2006-01-15

    Articular cartilage in adults has a limited capacity for self-repair after a substantial injury. Surgical therapeutic efforts to treat cartilage defects have focused on delivering new cells capable of chondrogenesis into the lesions. Autologous chondrocyte transplantation (ACT) is an advanced cell-based orthobiologic technology used for the treatment of chondral defects of the knee that has been in clinical use since 1987 and has been performed on 12,000 patients internationally. With ACT, good to excellent clinical results are seen in isolated post-traumatic lesions of the knee joint in the younger patient, with the formation of hyaline or hyaline-like repair tissue. In the classic ACT technique, chondrocytes are isolated from small slices of cartilage harvested arthroscopically from a minor weight-bearing area of the injured knee. The extracellular matrix is removed by enzymatic digestion, and the cells are then expanded in monolayer culture. Once a sufficient number of cells has been obtained, the chondrocytes are implanted into the cartilage defect, using a periosteal patch over the defect as a method of cell containment. The major complications are periosteal hypertrophy, delamination of the transplant, arthrofibrosis and transplant failure. Further improvements in tissue engineering have contributed to the next generation of ACT techniques, where cells are combined with resorbable biomaterials, as in matrix-associated autologous chondrocyte transplantation (MACT). These biomaterials secure the cells in the defect area and enhance their proliferation and differentiation.

  12. The Influence of Wealth and Race in Four-Year College Attendance

    OpenAIRE

    Su Jin Jez

    2008-01-01

    College is increasingly essential for economic and social mobility. Current research devotes significant attention to race and socioeconomic factors in college access. Yet wealth’s role, as differentiated from income, is largely unexplored. Utilizing a nationally representative dataset, this study analyzes the role of wealth among students who attend four-year colleges. The hypothesis that wealth matters through the provision of differential habitus, social capital, and cultural capital tha...

  13. Four-year-BS-position-paper.pdf | misc docs | academy | Indian ...

    Indian Academy of Sciences (India)

    Home; academy; misc docs; Four-year-BS-position-paper.pdf. 404! error. The page your are looking for can not be found! Please check the link or use the navigation bar at the top. YouTube; Twitter; Facebook; Blog. Academy News. IAS Logo. 29th Mid-year meeting. Posted on 19 January 2018. The 29th Mid-year meeting of ...

  14. Post-fire hydrologic response in Central Portugal. A four years study at microplot scale.

    Science.gov (United States)

    Vieira, Diana; Malvar, Maruxa; Martins, Martinho; Machado, Ana; Nunes, João; Keizer, Jacob

    2014-05-01

    Wildfires are a natural phenomenon in regions with a Mediterranean-type climate. However, their present-day widespread occurrence in southern Europe is unprecedented and strongly reflects human activity such as ignition, land-use changes, land abandonment and introduction of highly flammable plantations. Besides wildfires, post-fire management practices such as plowing, terracing, clearcutting and logging should also be considered, since their occurrence is getting increasingly common. And, in a long-term period these practices seem to be executed intercalated with repeated fire occurrences in the same site, sharing the impacts together with fire in an escalated degradational effect. In this sense, the work presented here concerns four years of runoff and erosion data at microplot scale after the wildfire, comparing different land management practices that occurred before the fire. Preliminary results indicate that in four years of monitoring, runoff is constantly higher in plowed sites than in the unplowed ones, with the exception of the first year. Regarding soil losses the plowed plots present always higher sediment rates than the unplowed ones. The comparison between two unplowed sites with different land uses, indicate higher runoff and erosion risk for pine comparatively to the eucalypt ones, however the reduced soil depth in the first can have an important role in these differences. Following these facts, the aim of the present work is to answer the following research questions: i) Do these four years of observations fit with the window of disturbance model presented by Prosser and Williams (1998). or the alternative version by Wittenberg and Inbar (2009)?; ii) Does pre-fire disturbances (wildfire, land use changes and land management practices) still have repercussions after wildfire?; In what sense does four years of intensive monitoring provides that one year couldn't provide?

  15. Habitual active transport, TV viewing and weight gain: a four year follow-up study.

    Science.gov (United States)

    Ding, Ding; Sugiyama, Takemi; Owen, Neville

    2012-01-01

    To examine the associations of TV viewing time and domain-specific physical activity with weight change; to determine whether domain-specific physical activity moderates the potential association of TV viewing time with weight change. We used four-year longitudinal data (baseline: 2003-2004, follow-up: 2007-2008) on 969 adults from selected neighborhoods in Adelaide, Australia (Age: 48.6 ± 10.6 years, 61% females). Mixed models examined four-year weight change as the dependent variable, with TV viewing time, habitual transport and past week domain-specific physical activity at baseline as independent variables. On average, participants gained 1.6 kg over four years. TV viewing time at baseline was positively associated with weight gain at follow-up. Each additional hour of TV viewing was associated with 0.24-0.27 kg of extra weight gain. This relationship was not moderated by recent recall of transport, leisure-time, and occupational physical activity, but was moderated by habitual transport: an additional hour of TV viewing time at baseline was significantly associated with an extra weight gain of 0.65 kg at follow-up among those who were inactive in everyday transport; TV time was not significantly associated with weight change among those who were regularly active in transport. Habitual active transport may protect adults against risk of weight gain associated with prolonged TV viewing time. Copyright © 2012 Elsevier Inc. All rights reserved.

  16. LOCAL INTERSTELLAR HYDROGEN'S DISAPPEARANCE AT 1 AU: FOUR YEARS OF IBEX IN THE RISING SOLAR CYCLE

    International Nuclear Information System (INIS)

    Saul, Lukas; Rodríguez, Diego; Scheer, Juergen; Wurz, Peter; Bzowski, Maciej; Kubiak, Marzena; Sokół, Justina; Fuselier, Stephen; McComas, Dave; Möbius, Eberhard

    2013-01-01

    NASA's Interstellar Boundary Explorer (IBEX) mission has recently opened a new window on the interstellar medium (ISM) by imaging neutral atoms. One ''bright'' feature in the sky is the interstellar wind flowing into the solar system. Composed of remnants of stellar explosions as well as primordial gas and plasma, the ISM is by no means uniform. The interaction of the local ISM with the solar wind shapes our heliospheric environment with hydrogen being the dominant component of the very local ISM. In this paper, we report on direct sampling of the neutral hydrogen of the local ISM over four years of IBEX observations. The hydrogen wind observed at 1 AU has decreased and nearly disappeared as the solar activity has increased over the last four years; the signal at 1 AU has dropped off in 2012 by a factor of ∼8 to near background levels. The longitudinal offset has also increased with time presumably due to greater radiation pressure deflecting the interstellar wind. We present longitudinal and latitudinal arrival direction measurements of the bulk flow as measured over four years beginning at near solar minimum conditions. The H distribution we observe at 1 AU is expected to be different from that outside the heliopause due to ionization, photon pressure, gravity, and filtration by interactions with heliospheric plasma populations. These observations provide an important benchmark for modeling of the global heliospheric interaction. Based on these observations we suggest a further course of scientific action to observe neutral hydrogen over a full solar cycle with IBEX.

  17. Stromal cell-derived factor 1 regulates the actin organization of chondrocytes and chondrocyte hypertrophy.

    Directory of Open Access Journals (Sweden)

    Koichi Murata

    Full Text Available Stromal cell-derived factor 1 (SDF-1/CXCL12/PBSF plays important roles in the biological and physiological functions of haematopoietic and mesenchymal stem cells. This chemokine regulates the formation of multiple organ systems during embryogenesis. However, its roles in skeletal development remain unclear. Here we investigated the roles of SDF-1 in chondrocyte differentiation. We demonstrated that SDF-1 protein was expressed at pre-hypertrophic and hypertrophic chondrocytes in the newly formed endochondral callus of rib fracture as well as in the growth plate of normal mouse tibia by immunohistochemical analysis. Using SDF-1(-/- mouse embryo, we histologically showed that the total length of the whole humeri of SDF-1(-/- mice was significantly shorter than that of wild-type mice, which was contributed mainly by shorter hypertrophic and calcified zones in SDF-1(-/- mice. Actin cytoskeleton of hypertrophic chondrocytes in SDF-1(-/- mouse humeri showed less F-actin and rounder shape than that of wild-type mice. Primary chondrocytes from SDF-1(-/- mice showed the enhanced formation of philopodia and loss of F-actin. The administration of SDF-1 to primary chondrocytes of wild-type mice and SDF-1(-/- mice promoted the formation of actin stress fibers. Organ culture of embryonic metatarsals from SDF-1(-/- mice showed the growth delay, which was recovered by an exogenous administration of SDF-1. mRNA expression of type X collagen in metatarsals and in primary chondrocytes of SDF-1(-/- mouse embryo was down-regulated while the administration of SDF-1 to metatarsals recovered. These data suggests that SDF-1 regulates the actin organization and stimulates bone growth by mediating chondrocyte hypertrophy.

  18. Stromal cell-derived factor 1 regulates the actin organization of chondrocytes and chondrocyte hypertrophy.

    Science.gov (United States)

    Murata, Koichi; Kitaori, Toshiyuki; Oishi, Shinya; Watanabe, Naoki; Yoshitomi, Hiroyuki; Tanida, Shimei; Ishikawa, Masahiro; Kasahara, Takashi; Shibuya, Hideyuki; Fujii, Nobutaka; Nagasawa, Takashi; Nakamura, Takashi; Ito, Hiromu

    2012-01-01

    Stromal cell-derived factor 1 (SDF-1/CXCL12/PBSF) plays important roles in the biological and physiological functions of haematopoietic and mesenchymal stem cells. This chemokine regulates the formation of multiple organ systems during embryogenesis. However, its roles in skeletal development remain unclear. Here we investigated the roles of SDF-1 in chondrocyte differentiation. We demonstrated that SDF-1 protein was expressed at pre-hypertrophic and hypertrophic chondrocytes in the newly formed endochondral callus of rib fracture as well as in the growth plate of normal mouse tibia by immunohistochemical analysis. Using SDF-1(-/-) mouse embryo, we histologically showed that the total length of the whole humeri of SDF-1(-/-) mice was significantly shorter than that of wild-type mice, which was contributed mainly by shorter hypertrophic and calcified zones in SDF-1(-/-) mice. Actin cytoskeleton of hypertrophic chondrocytes in SDF-1(-/-) mouse humeri showed less F-actin and rounder shape than that of wild-type mice. Primary chondrocytes from SDF-1(-/-) mice showed the enhanced formation of philopodia and loss of F-actin. The administration of SDF-1 to primary chondrocytes of wild-type mice and SDF-1(-/-) mice promoted the formation of actin stress fibers. Organ culture of embryonic metatarsals from SDF-1(-/-) mice showed the growth delay, which was recovered by an exogenous administration of SDF-1. mRNA expression of type X collagen in metatarsals and in primary chondrocytes of SDF-1(-/-) mouse embryo was down-regulated while the administration of SDF-1 to metatarsals recovered. These data suggests that SDF-1 regulates the actin organization and stimulates bone growth by mediating chondrocyte hypertrophy.

  19. Genetic modification of chondrocytes with insulin-like growth factor-1 enhances cartilage healing in an equine model.

    Science.gov (United States)

    Goodrich, L R; Hidaka, C; Robbins, P D; Evans, C H; Nixon, A J

    2007-05-01

    Gene therapy with insulin-like growth factor-1 (IGF-1) increases matrix production and enhances chondrocyte proliferation and survival in vitro. The purpose of this study was to determine whether arthroscopically-grafted chondrocytes genetically modified by an adenovirus vector encoding equine IGF-1 (AdIGF-1) would have a beneficial effect on cartilage healing in an equine femoropatellar joint model. A total of 16 horses underwent arthroscopic repair of a single 15 mm cartilage defect in each femoropatellar joint. One joint received 2 x 10(7) AdIGF-1 modified chondrocytes and the contralateral joint received 2 x 10(7) naive (unmodified) chondrocytes. Repairs were analysed at four weeks, nine weeks and eight months after surgery. Morphological and histological appearance, IGF-1 and collagen type II gene expression (polymerase chain reaction, in situ hybridisation and immunohistochemistry), collagen type II content (cyanogen bromide and sodium dodecyl sulphate-polyacrylamide gel electrophoresis), proteoglycan content (dimethylmethylene blue assay), and gene expression for collagen type I, matrix metalloproteinase (MMP)-1, MMP-3, MMP-13, aggrecanase-1, tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) and TIMP-3 were evaluated. Genetic modification of chondrocytes significantly increased IGF-1 mRNA and ligand production in repair tissue for up to nine weeks following transplantation. The gross and histological appearance of IGF-1 modified repair tissue was improved over control defects. Gross filling of defects was significantly improved at four weeks, and a more hyaline-like tissue covered the lesions at eight months. Histological outcome at four and nine weeks post-transplantation revealed greater tissue filling of defects transplanted with genetically modified chondrocytes, whereas repair tissue in control defects was thin and irregular and more fibrous. Collagen type II expression in IGF-1 gene-transduced defects was increased 100-fold at four weeks and

  20. Three-year clinical outcome after chondrocyte transplantation using a hyaluronan matrix for cartilage repair

    Energy Technology Data Exchange (ETDEWEB)

    Nehrer, S. [Department of Orthopedics, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna (Austria)]. E-mail: stefan.nehrer@meduniwien.ac.at; Domayer, S. [Department of Orthopedics, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna (Austria); Dorotka, R. [Department of Orthopedics, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna (Austria); Schatz, K. [Department of Orthopedics, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna (Austria); Bindreiter, U. [Department of Orthopedics, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna (Austria); Kotz, R. [Department of Orthopedics, Medical University of Vienna, Waehringer Guertel 18-20, A-1090 Vienna (Austria)

    2006-01-15

    Repair of articular cartilage represents a significant clinical problem and although various new techniques - including the use of autologous chondrocytes - have been developed within the last century the clinical efficacy of these procedures is still discussed controversially. Although autologous chondrocyte transplantation (ACT) has been widely used with success, it has several inherent limitations, including its invasive nature and problems related to the use of the periosteal flap. To overcome these problems autologous chondrocytes transplantation combined with the use of biodegradable scaffolds has received wide attention. Among these, a hyaluronan-based scaffold has been found useful for inducing hyaline cartilage regeneration. In the present study, we have investigated the mid-term efficacy and safety of Hyalograft[reg] C grafts in a group of 36 patients undergoing surgery for chronic cartilage lesions of the knee. Clinical Outcome was assessed prospectively before and at 12, 24, and 36 months after surgery. No major adverse events have been reported during the 3-year follow-up. Significant improvements of the evaluated scores were observed (P < 0.02) at 1 year and a continued increase of clinical performance was evident at 2 and 3 years follow-up. Patients under 30 years of age with single lesions showed statistically significant improvements at all follow-up visits compared to those over 30 with multiple defects (P < 0.01). Hyalograft[reg] C compares favorably with classic ACT and is particularly indicated in younger patients with single lesions. The graft can be implanted through a miniarthrotomy and needs no additional fixation with sutures except optional fibrin gluing at the defect borders. These results suggest that Hyalograft[reg] C is a valid alternative to ACT.

  1. Pancreas grafts

    International Nuclear Information System (INIS)

    Hahn, D.; Buell, U.; Land, W.; Unertl, K.

    1981-01-01

    Perfusion studies with sup(99m)Tc-DTPA, which has hitherto been used routinely to investigate renal grafts, have also proved useful for monitoring the perfusion of pancreas grafts. A total perfusion failure is equally reliably demonstrable as in renal grafts. Quantitatively smaller perfusion alterations can be demonstrated by monitoring the course. It seems possible to differentiate the salivary edema of a rejection reaction, well known from animal experiments, with the help of other paramters (e.g. creatinine). Further clinical studies are however necessary to confirm these results. (orig.) [de

  2. Dermatitis and lymphadenitis resembling juvenile cellulitis in a four-year-old dog.

    Science.gov (United States)

    Neuber, A E; van den Broek, A H M; Brownstein, D; Thoday, K L; Hill, P B

    2004-05-01

    A four-year-old, entire male toy poodle was presented with a two-and-a-half-week history of ocular discharge progressing to periorbital alopecia, depigmentation, alopecia and ulceration around the muzzle. There was also a haemorrhagic discharge from the ears, pyrexia, lethargy and generalised lymphadenopathy. The clinical, cytological, bacteriological and histopathological findings were consistent with a diagnosis of dermatitis resembling juvenile cellulitis in an adult dog. Glucocorticoid therapy led to rapid resolution of the clinical signs and the dog has remained in remission for two years after cessation of treatment.

  3. Birth centre confinement at the Queen Victoria Medical Centre: four years' experience.

    Science.gov (United States)

    Morris, N; Campbell, J; Biro, M A; Lumley, J; Rao, J; Spensley, J

    1986-06-09

    A review of the first four years of the functioning of the birth centre at the Queen Victoria Medical Centre is presented. In that time, 1040 pregnant women were accepted for confinement there. Of these, 52 withdrew for non-obstetric reasons, while 470 were transferred to alternative obstetrical care--274 because of antepartum complications and 196 because of intrapartum problems. Therefore, 518 women were delivered in the birth centre. The care of the women is entrusted almost entirely to a team of midwives and this review demonstrates an enviable safety record.

  4. Report on the green paper on energy. Four years of European initiatives

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2005-07-01

    In December 2000, the European Commission adopted a Green Paper on European Union energy policy. This brochure takes stock of the action undertaken in the following areas over the last four years:managing demand, diversifying internal energy sources,developing the internal energy market and the security of external supply. It presents: the Green Paper stakes, the progress made, the four political challenges, managing demand, diversifying European sources, the streamlined internal energy market, controlling external supply, future prospects for the Union, legislative developments and ten possibilities for economical energy use. (A.L.B.)

  5. A four-year surveillance program for detection of Plasmodium falciparum chloroquine resistance in Honduras

    Directory of Open Access Journals (Sweden)

    Gustavo A Fontecha

    2014-07-01

    Full Text Available Countries could use the monitoring of drug resistance in malaria parasites as an effective early warning system to develop the timely response mechanisms that are required to avert the further spread of malaria. Drug resistance surveillance is essential in areas where no drug resistance has been reported, especially if neighbouring countries have previously reported resistance. Here, we present the results of a four-year surveillance program based on the sequencing of the pfcrt gene of Plasmodium falciparum populations from endemic areas of Honduras. All isolates were susceptible to chloroquine, as revealed by the pfcrt “CVMNK” genotype in codons 72-76.

  6. A four-year surveillance program for detection of Plasmodium falciparum chloroquine resistance in Honduras.

    Science.gov (United States)

    Fontecha, Gustavo A; Sanchez, Ana L; Mendoza, Meisy; Banegas, Engels; Mejía-Torres, Rosa E

    2014-07-01

    Countries could use the monitoring of drug resistance in malaria parasites as an effective early warning system to develop the timely response mechanisms that are required to avert the further spread of malaria. Drug resistance surveillance is essential in areas where no drug resistance has been reported, especially if neighbouring countries have previously reported resistance. Here, we present the results of a four-year surveillance program based on the sequencing of the pfcrt gene of Plasmodium falciparum populations from endemic areas of Honduras. All isolates were susceptible to chloroquine, as revealed by the pfcrt "CVMNK" genotype in codons 72-76.

  7. Four Years of Real-Time GRB Followup by BOOTES-1B (2005–2008

    Directory of Open Access Journals (Sweden)

    Martin Jelínek

    2010-01-01

    Full Text Available Four years of BOOTES-1B GRB follow-up history are summarised for the first time in the form of a table. The successfully followed events are described case by case. Further, the data are used to show the GRB trigger rate in Spain on a per-year basis, resulting in an estimate of 18 triggers and about 51 hours of telescope time per year for real-time triggers. These numbers grow to about 22 triggers and 77 hours per year if we include also the GRBs observable within 2 hours after the trigger.

  8. Report on the green paper on energy. Four years of European initiatives

    International Nuclear Information System (INIS)

    2005-01-01

    In December 2000, the European Commission adopted a Green Paper on European Union energy policy. This brochure takes stock of the action undertaken in the following areas over the last four years:managing demand, diversifying internal energy sources,developing the internal energy market and the security of external supply. It presents: the Green Paper stakes, the progress made, the four political challenges, managing demand, diversifying European sources, the streamlined internal energy market, controlling external supply, future prospects for the Union, legislative developments and ten possibilities for economical energy use. (A.L.B.)

  9. A FOUR-YEAR CHRONOLOGY WITH NATIONAL TEAM BOXING IN CANADA

    Directory of Open Access Journals (Sweden)

    Robert J. Schinke

    2007-10-01

    Full Text Available Applied sport psychologists tend to begin their consulting relationships with national teams having formalized skills, and often, limited contextual and sport- specific understanding. The present report overviews the first four years of a long-term consulting relationship one practitioner developed with the Canadian National Boxing Team. From the vantage of an applied sport psychology consultant, I overview how a limited consulting role expanded into increased responsibilities and opportunities. Suggestions are provided for the aspiring sport psychology consultant interested in working with athletes and coaches within the combative sport of elite amateur boxing

  10. Describing medical student curiosity across a four year curriculum: An exploratory study.

    Science.gov (United States)

    Sternszus, Robert; Saroyan, Alenoush; Steinert, Yvonne

    2017-04-01

    Intellectual curiosity can be defined as a desire for knowledge that leads to exploratory behavior and consists of an inherent and stable trait (i.e. trait curiosity) and a variable context-dependent state (i.e. state curiosity). Although intellectual curiosity has been considered an important aspect of medical education and practice, its relationship to medical education has not been empirically investigated. The purpose of this exploratory study was to describe medical students' intellectual curiosity across a four-year undergraduate program. We employed a cross-sectional design in which medical students, across a four-year undergraduate program at McGill University, completed the Melbourne Curiosity Inventory as a measure of their state and trait intellectual curiosity. A Mixed Models ANOVA was used to compare students across year of training. Four hundred and two out of 751 students completed the inventory (53.5%). Trait curiosity was significantly higher than state curiosity (M = 64.5, SD = 8.5 versus M = 58.5, SD = 11.6) overall, and within each year of training. This study is the first to describe state and trait intellectual curiosity in undergraduate medical education. Findings suggest that medical students' state curiosity may not be optimally supported and highlight avenues for further research.

  11. Evolution of upper limb kinematics four years after subacute robot-assisted rehabilitation in stroke patients.

    Science.gov (United States)

    Pila, Ophélie; Duret, Christophe; Gracies, Jean-Michel; Francisco, Gerard E; Bayle, Nicolas; Hutin, Émilie

    2018-04-25

    To assess functional status and robot-based kinematic measures four years after subacute robot-assisted rehabilitation in hemiparesis. Twenty-two patients with stroke-induced hemiparesis underwent a ≥3-month upper limb combined program of robot-assisted and occupational therapy from two months post-stroke, and received community-based therapy after discharge. Four years later, 19 (86%) participated in this follow-up study. Assessments 2, 5 and 54 months post-stroke included Fugl-Meyer (FM), Modified Frenchay Scale (MFS, at Month 54) and robot-based kinematic measures of targeting tasks in three directions, north, paretic and non-paretic: distance covered, velocity, accuracy (root mean square (RMS) error from straight line) and smoothness (number of velocity peaks; upward changes in accuracy and smoothness represent worsening). Analysis was stratified by FM score at two months: ≥17 (Group 1) or Kinematic changes (three directions pooled) were: distance -1[-17;2]% (ns); velocity, -8[-32;28]% (ns); accuracy, +6[-13;98]% (ns); smoothness, +44[-6;126]% (p robot-assisted upper limb training during subacute post-stroke phase, movement kinematics deteriorated despite community-based therapy, especially in more severely impaired patients. EudraCT 2016-005121-36. Registration: 2016-12-20. Date of enrolment of the first participant to the trial: 2009-11-24.

  12. Dutch healthcare professionals inadequately perceived if three- and four-year-old preschool children were overweight.

    Science.gov (United States)

    Bocca, Gianni; Corpeleijn, Eva; Broens, Jasper; Stolk, Ronald P; Sauer, Pieter J

    2016-10-01

    We studied whether healthcare professionals adequately perceived if preschool children were overweight and whether this was influenced by their own body mass index (BMI). We sent 716 Dutch healthcare professionals questionnaires containing seven pictures and seven sketches of three- and four-year-old children showing body weights from underweight to morbidly obese. The professionals rated the pictures on a five-point scale from too heavy to too light and chose the sketch that they felt best depicted the child's body shape. They also reported their own height and weight and their BMI was calculated. Of the 716 questionnaires, 346 (48.3%) were returned with complete information and analysed. Healthcare professionals mostly chose sketches that showed children as being lighter than they really were. Depending on their own BMI group, the overweight child was perceived as having a normal weight by 74-79% of the healthcare professionals. The obese children were rated correctly by 44-52% of the professionals, but 14-15% said their weight was normal. The morbidly obese child was adequately assessed by 93-98% of the professionals. Healthcare professionals inadequately perceived whether three- and four-year-old children were overweight and this may have hindered early interventions, leading to overweight children becoming overweight adolescents. ©2015 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.

  13. Increasing resistance to quinolones: A four-year prospective study of urinary tract infection pathogens

    Directory of Open Access Journals (Sweden)

    Orhiosefe Omigie

    2009-08-01

    Full Text Available Orhiosefe Omigie, Lawrence Okoror, Patience Umolu, Gladys IkuuhDepartment of Microbiology, Ambrose Alli University, Ekpoma, NigeriaAbstract: A four-year prospective study was carried out to determine the incidence and rate of development of resistance by common urinary tract infection (UTI pathogens to quinolone antimicrobial agents. Results show that there is high intrinsic resistance to the quinolones among strains of Pseudomonas aeruginosa (43.4%, Escherichia coli (26.3%, and Proteus spp. (17.1%. Over four years, rising rates of resistance were observed in P. aeruginosa (14.6% increase, Staphylococcus aureus (9.8%, and E. coli (9.7%. The highest potency was exhibited by ciprofloxacin (91.2%, levofloxacin (89.2%, and moxifloxacin (85.1%, while there were high rates of resistance to nalidixic acid (51.7% and pefloxacin (29.0%. Coliforms, particularly E. coli (>45%, remain the most prevalent causative agents of UTI while females within the age range of 20–50 years were most vulnerable to UTI.Keywords: UTI, microorganisms, antibiotics, resistance

  14. IPS Empress inlays and onlays after four years--a clinical study.

    Science.gov (United States)

    Krämer, N; Frankenberger, R; Pelka, M; Petschelt, A

    1999-07-01

    Ceramic inlays are used as esthetic alternatives to amalgam and other metallic materials for the restoration of badly damaged teeth. However, only limited clinical data are available regarding adhesive inlays and onlays with proximal margins located in dentine. In a prospective, controlled clinical study, the performance of IPS Empress inlays and onlays with cuspal replacements and margins below the amelocemental junction was examined. Ninety-six IPS Empress fillings were placed in 34 patients by six clinicians. The restorations were luted with four different composite systems. The dentin bonding system Syntac Classic was used in addition to the acid-etch-technique. At baseline and after 6 months, one, two and four years after placement the restorations were assessed by two calibrated investigators using modified USPHS codes and criteria. A representative sample of the restorations was investigated by scanning electron microscopy to evaluate wear. Seven of the 96 restorations investigated had to be replaced (failure rate 7%; Kaplan-Meier). Four inlays had suffered cohesive bulk fractures and three teeth required endodontic treatment. After four years in clinical service, significant deterioration (Friedman 2-way Anova; p Empress inlays and onlays bonded with the dentin bonding system Syntac Classic were found to have a 7% failure rate with 79% of the remaining restorations having marginal deficiencies.

  15. Overweight among Four-Year-Old Children in Relation to Early Growth Characteristics and Socioeconomic Factors

    Directory of Open Access Journals (Sweden)

    Jörgen Thorn

    2010-01-01

    Full Text Available Objectives. To assess early growth characteristics and socioeconomic factors of children in relation to body mass index (BMI and presence of overweight among four-year-old children. Methods. Two Child Health Centres (CHC participated in the study. They were selected to obtain two populations of children featuring divergent socio-economic characteristics. Growth data registered at the CHCs from birth to the 4-year check-up were recovered. Overweight was defined by the BMI cut-offs established by IOTF. BMI values expressed as BMI standard deviation score (BMISDS were used for analysis. Results. At the 4-year check-up, the BMISDS and the proportion of children with overweight (including the obese were significantly higher in the district with lower socio-economic status. High BMI at birth and low socio-economic status of the population in the CHC-district were shown to be independent determinants for overweight and BMISDS at four years of age. Conclusions. More research is needed to understand the mechanisms and how intervention programs should be designed in order to prevent the development of overweight and obesity in children.

  16. An evaluation of vocational high schools in Indonesia: A comparison between four-year and three-year programs

    Directory of Open Access Journals (Sweden)

    Soenarto Soenarto

    2017-12-01

    Full Text Available The research aimed to gain insights into the quality of four-year program vocational high school (VHS in Indonesia compared to four-year program VHS. This research was conducted based on the school graduate standard, business sector and industrial sector (or Dunia Usaha dan Dunia Industri (DUDI – or the performance of the graduates and alumni (the graduates’ satisfaction. The research was conducted using Discrepancy Evaluation Model using 16 VHSs (eight four-year program VHSs and eight three-year program VHSs. The result shows that from the standpoint of the school, the graduates of the four-year program VHS are higher in quality than those of the three-year program VHS. The four-year program VHS graduates are more qualified in seven aspects: teamwork, discipline, tenacity, theoretical knowledge, confidence, creativity, and leadership. Meanwhile, using DUDI standpoint, the four-year program VHS graduates are also higher in quality than the three-year program VHS graduates. In addition, the four-year program VHS graduates are better in the quality of their discipline, tenacity, theoretical knowledge, practical skills, confidence, carefulness, creativity, and leadership. The four-year program VHS graduates have a higher level of satisfaction in terms of income than the three-year program VHS graduates. The higher quality of the four-year program VHS graduates has resulted from longer duration of the internship program (PKL that provides them with reliable experience and skills concerning work-related problem-solving activities.

  17. Experimental study of tissue-engineered cartilage allograft with RNAi chondrocytes in vivo

    Directory of Open Access Journals (Sweden)

    Wang ZH

    2014-05-01

    Full Text Available Zhenghui Wang,1 Xiaoli Li,2 Xi-Jing He,3 Xianghong Zhang,1 Zhuangqun Yang,4 Min Xu,1 Baojun Wu,1 Junbo Tu,5 Huanan Luo,1 Jing Yan11Department of Otolaryngology – Head and Neck Surgery, 2Department of Dermatology, 3Department of Orthopedics, The Second Hospital, Xi’an Jiaotong University, 4Department of Plastic and Burns Surgery, The First Hospital, Xi’an Jiaotong University, 5Department of Oral and Maxillofacial Plastic Surgery, The Stomatological Hospital, Xi’an Jiaotong University, Xi’an, People’s Republic of ChinaPurpose: To determine the effects of RNA interference (RNAi on chondrocyte proliferation, function, and immunological rejection after allogenic tissue-engineered cartilage transplantation within bone matrix gelatin scaffolds.Methods: Seven million rat normal and RNAi chondrocytes were harvested and separately composited with fibrin glue to make the cell suspension, and then transplanted subcutaneously into the back of Sprague Dawley rats after being cultured for 10 days in vitro. Untransplanted animals served as the control group. The allograft and immunological response were examined at 1, 2, 4, 8, and 12 months postoperatively with hematoxylin and eosin histochemical staining, immunohistochemical staining (aggrecan, type II collagen, class I and II major histocompatibility complex, and flow cytometry for peripheral blood cluster of differentiation 4+ (CD4+ and CD8+ T-cells.Results: There was no infection or death in the rats except one, which died in the first week. Compared to the control group, the RNAi group had fewer eukomonocytes infiltrated, which were only distributed around the graft. The ratio of CD4+/CD8+ T-cells in the RNAi group was significantly lower than the normal one (P<0.05. There were many more positively stained chondrocytes and positively stained areas around the cells in the RNAi group, which were not found in the control group.Conclusion: The aggrecanase-1 and aggrecanase-2 RNAi for chondrocytes

  18. EVOLUTION OF THE ROMANIAN CAPITAL MARKET IN THE LAST FOUR YEARS

    Directory of Open Access Journals (Sweden)

    SORIN CLAUDIU RADU

    2012-10-01

    Full Text Available The effects of the great recession have also been felt in Romania and the domestic capital market, component of the European financial market, has suffered from the negative evolutions of the world’s economy, fallowing the downward trend of the external markets since the crisis started. Considering the economic situation as of 2008, the Romanian capital market was faced with a small number of transactions, a decreased stock capitalization and low level of credibility, massive diminishment of liquidities, capital withdrawal on the financial market and significant depreciation of stock exchange indicators. This paper aims at presenting the evolution of the capital market in Romania for the last four years, pinpointing several strategic directions that are meant to support the domestic stock exchange and place them in the European stock exchanges top.

  19. Paediatric Stroke: A Rare Presentation of Iron Deficiency Anemia in a Four Year Old Child.

    Science.gov (United States)

    Subedi, K; Koirala, S; Basnet, R; Poudel, A

    Paediatric stroke is an uncommon syndrome with even lesser annual incidence rates of arterial ischemic stroke in infants and children. In children the diagnosis of stroke is frequently delayed or missed. This is due to subtle and nonspecific clinical presentations, a complicated differential diagnosis and a lack of awareness by physicians and also delay in the seeking of medical attention as in our case. We report you a rare case of a four year old child from remote Nepal who presented to our Out Patient Department after a long gap of around four months after the sudden onset of loss of consciousness and decreased movement of right limbs who after detailed history examination and lab investigations and imaging revealed ischemic stroke due to iron deficiency anemia.

  20. The Nuclear Organization and Management Analysis Concept methodology: Four years later

    International Nuclear Information System (INIS)

    Haber, S.B.; Shurberg, D.A.; Barriere, M.T.; Hall, R.E.

    1992-01-01

    The Nuclear Organization and Management Analysis Concept was first presented at the IEEE Human Factors meeting in Monterey in 1988. In the four years since that paper, the concept and its associated methodology has been demonstrated at two commercial nuclear power plants (NPP) and one fossil power plant. In addition, applications of some of the methods have been utilized in other types of organizations, and products are being developed from the insights obtained using the concept for various organization and management activities. This paper will focus on the insights and results obtained from the two demonstration studies at the commercial NPPs. The results emphasize the utility of the methodology and the comparability of the results from the two organizations

  1. Cerebral vein thrombosis in a four year old with Behçet's disease.

    Science.gov (United States)

    Hacihamdioglu, Duygu Ovunc; Demiriz, Murat; Sobaci, Gungor; Kocaoglu, Murat; Demirkaya, Erkan; Gok, Faysal

    2014-01-01

    Behçet's disease (BD) is a multisystem disorder. The main pathology in BD is vasculitis that involves arteries and veins of all calibers. Central nervous system involvement occurs in 5-10% of patients. Increased morbidity and mortality is rarely observed in children. The mean age at onset in pediatric BD is approximately 7 years. Neurologic involvement in BD is usually observed after 3-6 years. We report the case of a four-year-old Turkish boy with BD with sagittal sinus thrombosis treated with infliximab. The patient presented papilledema without neurologic signs. Although long-term efficacy evaluations are needed in this case, infliximab therapy may be a good option in childhood BD with refractory sinus thrombosis. This is the youngest case of BD with sagittal sinus thrombosis reported so far. Copyright © 2012 Elsevier España, S.L. All rights reserved.

  2. Rotan manau intercropped with rubber: rate of root growth between three and four years after planting

    International Nuclear Information System (INIS)

    Wan Rashidah Kadir; Aminuddin Mohamad; Ahmad Sahali Mardi; Zaharah Abd Rahman

    1997-01-01

    Wan Rashidah, K., Aminuddin, M., Ahmad Sahali, M. & Zaharah, A.R. 1997. Rotan manau intercropped with rubber: rate of root growth between three and four years after planting. Efficient fertiliser management depends partly on understanding the active root distribution. In the present study, the active root distribution of 3- and 4-y-old plantation grown rotan manau (Calamus manan) was assessed using isotope tracer technique. For the 3-y-old rotan manau, three distances from the plant base (0.5, 1.0 and 1.5 m) at 5 and 30 cm depths were examined. For the 4-y-old plants, two distances, viz-a-viz at a centre between two rattan plants and another in the middle between two rattan plants and two rubber trees were studied. The isotope used was 32P, applied as a solution with KH2PO4. The rotan manau plants had been established under mature rubber plantation. High proportions of feeder roots were found at 0.5 and 1.0 m distances at the surface (5 cm depth) for the 3-y-old plants. Uptake of 32P was also observed for the application at 1.5 m distance for both depths but the counts were small. Statistical analysis gave a highly significant difference within the distances and within the different depths. For a better synchronisation between fertiliser application and plant uptake, it seems that application at approximately between 0.5 and 1.0 m distance around the plant is most appropriate at this age. At four years after planting, important uptake was obtained only for the two plants located near the application area. Anyhow, to some extent it reflected that roots had already extended for another 1 m compared to the 3-y-old plants

  3. A cause of Sudden Cardiac Deaths on Autopsy Findings; a Four-Year Report.

    Science.gov (United States)

    Rao, Dinesh; Sood, Divya; Pathak, P; Dongre, Sudhir D

    2014-01-01

    Incidence of sudden cardiac death (SCD) has been steadily increasing all over the world. While knowing the cause of SCD is one of the favorites of the physicians involved with these cases, it is very difficult and challenging task for the forensic physician. The present report is a prospective study regarding cause of SCDs on autopsy examination in four-year period, Bangalore, India. The present prospective study is based on autopsy observations, carried out for four-year period from 2008 to 2011, and analyzed for cause of SCDs. The cases were chosen as per the definition of sudden death and autopsied. The material was divided into natural and unnatural groups. Finally, on histopathology, gross examination, hospital details, circumstantial, and police reports the cause of death was inferred. A total of 2449 autopsy was conducted of which 204 cases were due to SCD. The highest SCDs were reported in 50-60 years age group (62.24%; n-127), followed closely by the age group 60-69 (28.43%; n-58). Male to female ratio was around 10:1. The maximum number of deaths (n=78) was within few hours (6 hours) after the onset of signs and symptoms. In 24 (11.8%) cases major narrowing was noted in both the main coronaries, in 87 (42.6%) cases in the left anterior descending coronary artery (LAD), and in 18 (51.5%) cases in the right coronary artery (RCA). The major cardiac pathology resulting in sudden death was coronary artery disease (n-116; 56.86%) and myocardial infarction (n-104; 50.9%). most of the SCDs occurred in the place of residence (n-80; 39.2%) followed closely by death in hospital (n-49; 24.01%). Coronary occlusion was the major contributory cause of sudden death with cardiac origin and the highest number of deaths were reported in the age 50-59 years with male to female ratio of 10:1.

  4. Bobath or motor relearning programme? A follow-up one and four years post stroke.

    Science.gov (United States)

    Langhammer, Birgitta; Stanghelle, Johan K

    2003-11-01

    The purpose of this follow-up one and four years post stroke was to find out whether the initial physiotherapy approach had had any long-term effects on mortality, motor function, postural control, activities of daily living, life quality, follow-up from community services and living conditions. A randomized controlled trial of first time ever stroke patients. Group 1 (n = 33) and group 2 (n = 28) had initial physiotherapy according to the Motor Relearning Programme and Bobath, respectively. The Motor Assessment Scale (MAS), the Sødring Motor Evaluation Scale (SMES), the Barthel ADL Index, the Nottingham Health Profile (NHP) and Berg Balance Scale were used. The following parameters were also registered: incidence of new strokes, other diseases, use of assistive devices, the patient's accommodation and use of services from the community. The mortality rates were similar in the two groups. In both groups the motor function, postural control and ADL had decreased rapidly, leaving many of the patients dependent and with a high risk of falling. Life quality had increased compared to the acute stage, but was still low in comparison with healthy persons. Patients in both groups lived at home, but were dependent on help from relatives and community services. Physiotherapy as follow-up service was seldom used. The initial physiotherapy approach did not seem to have a major influence on the patients' ability to cope in the long-term. This follow-up at one and four years post stroke showed no major influence of two different initial physiotherapy regimens on long-term function. The study confirmed a rapid deterioration of ADL and motor function and an increased dependence on relatives. The study reveals a gap between the intense treatment in the acute phase and little or no follow-up of physiotherapy treatment or other rehabilitation activities later.

  5. Nasal chondrocyte-based engineered autologous cartilage tissue for repair of articular cartilage defects: an observational first-in-human trial.

    Science.gov (United States)

    Mumme, Marcus; Barbero, Andrea; Miot, Sylvie; Wixmerten, Anke; Feliciano, Sandra; Wolf, Francine; Asnaghi, Adelaide M; Baumhoer, Daniel; Bieri, Oliver; Kretzschmar, Martin; Pagenstert, Geert; Haug, Martin; Schaefer, Dirk J; Martin, Ivan; Jakob, Marcel

    2016-10-22

    Articular cartilage injuries have poor repair capacity, leading to progressive joint damage, and cannot be restored predictably by either conventional treatments or advanced therapies based on implantation of articular chondrocytes. Compared with articular chondrocytes, chondrocytes derived from the nasal septum have superior and more reproducible capacity to generate hyaline-like cartilage tissues, with the plasticity to adapt to a joint environment. We aimed to assess whether engineered autologous nasal chondrocyte-based cartilage grafts allow safe and functional restoration of knee cartilage defects. In a first-in-human trial, ten patients with symptomatic, post-traumatic, full-thickness cartilage lesions (2-6 cm 2 ) on the femoral condyle or trochlea were treated at University Hospital Basel in Switzerland. Chondrocytes isolated from a 6 mm nasal septum biopsy specimen were expanded and cultured onto collagen membranes to engineer cartilage grafts (30 × 40 × 2 mm). The engineered tissues were implanted into the femoral defects via mini-arthrotomy and assessed up to 24 months after surgery. Primary outcomes were feasibility and safety of the procedure. Secondary outcomes included self-assessed clinical scores and MRI-based estimation of morphological and compositional quality of the repair tissue. This study is registered with ClinicalTrials.gov, number NCT01605201. The study is ongoing, with an approved extension to 25 patients. For every patient, it was feasible to manufacture cartilaginous grafts with nasal chondrocytes embedded in an extracellular matrix rich in glycosaminoglycan and type II collagen. Engineered tissues were stable through handling with forceps and could be secured in the injured joints. No adverse reactions were recorded and self-assessed clinical scores for pain, knee function, and quality of life were improved significantly from before surgery to 24 months after surgery. Radiological assessments indicated variable degrees of

  6. Considerations on the use of ear chondrocytes as donor chondrocytes for cartilage tissue engineering

    NARCIS (Netherlands)

    van Osch, Gerjo J. V. M.; Mandl, Erik W.; Jahr, Holger; Koevoet, Wendy; Nolst-Trenité, Gilbert; Verhaar, Jan A. N.

    2004-01-01

    Articular cartilage is often used for research on cartilage tissue engineering. However, ear cartilage is easier to harvest, with less donor-site morbidity. The aim of this study was to evaluate whether adult human ear chondrocytes were capable of producing cartilage after expansion in monolayer

  7. Chondrocyte-specific ablation of Osterix leads to impaired endochondral ossification

    Energy Technology Data Exchange (ETDEWEB)

    Oh, Jung-Hoon [Department of Molecular Medicine, Cell and Matrix Research Institute, BK21 Medical Education Program for Human Resources, Kyungpook National University School of Medicine, Daegu (Korea, Republic of); Park, Seung-Yoon [Department of Biochemistry, School of Medicine, Dongguk University, Gyeongju 780-714 (Korea, Republic of); Crombrugghe, Benoit de [Department of Genetics, University of Texas, M.D. Anderson Cancer Center, Houston (United States); Kim, Jung-Eun, E-mail: kjeun@knu.ac.kr [Department of Molecular Medicine, Cell and Matrix Research Institute, BK21 Medical Education Program for Human Resources, Kyungpook National University School of Medicine, Daegu (Korea, Republic of)

    2012-02-24

    Highlights: Black-Right-Pointing-Pointer Conditional ablation of Osterix (Osx) in chondrocytes leads to skeletal defects. Black-Right-Pointing-Pointer Osx regulates chondrocyte differentiation and bone growth in growth plate chondrocytes. Black-Right-Pointing-Pointer Osx has an autonomous function in chondrocytes during endochondral ossification. -- Abstract: Osterix (Osx) is an essential transcription factor required for osteoblast differentiation during both intramembranous and endochondral ossification. Endochondral ossification, a process in which bone formation initiates from a cartilage intermediate, is crucial for skeletal development and growth. Osx is expressed in differentiating chondrocytes as well as osteoblasts during mouse development, but its role in chondrocytes has not been well studied. Here, the in vivo function of Osx in chondrocytes was examined in a chondrocyte-specific Osx conditional knockout model using Col2a1-Cre. Chondrocyte-specific Osx deficiency resulted in a weak and bent skeleton which was evident in newborn by radiographic analysis and skeletal preparation. To further understand the skeletal deformity of the chondrocyte-specific Osx conditional knockout, histological analysis was performed on developing long bones during embryogenesis. Hypertrophic chondrocytes were expanded, the formation of bone trabeculae and marrow cavities was remarkably delayed, and subsequent skeletal growth was reduced. The expression of several chondrocyte differentiation markers was reduced, indicating the impairment of chondrocyte differentiation and endochondral ossification in the chondrocyte-specific Osx conditional knockout. Taken together, Osx regulates chondrocyte differentiation and bone growth in growth plate chondrocytes, suggesting an autonomous function of Osx in chondrocytes during endochondral ossification.

  8. Applications of Chondrocyte-Based Cartilage Engineering: An Overview

    Directory of Open Access Journals (Sweden)

    Abdul-Rehman Phull

    2016-01-01

    Full Text Available Chondrocytes are the exclusive cells residing in cartilage and maintain the functionality of cartilage tissue. Series of biocomponents such as different growth factors, cytokines, and transcriptional factors regulate the mesenchymal stem cells (MSCs differentiation to chondrocytes. The number of chondrocytes and dedifferentiation are the key limitations in subsequent clinical application of the chondrocytes. Different culture methods are being developed to overcome such issues. Using tissue engineering and cell based approaches, chondrocytes offer prominent therapeutic option specifically in orthopedics for cartilage repair and to treat ailments such as tracheal defects, facial reconstruction, and urinary incontinence. Matrix-assisted autologous chondrocyte transplantation/implantation is an improved version of traditional autologous chondrocyte transplantation (ACT method. An increasing number of studies show the clinical significance of this technique for the chondral lesions treatment. Literature survey was carried out to address clinical and functional findings by using various ACT procedures. The current study was conducted to study the pharmacological significance and biomedical application of chondrocytes. Furthermore, it is inferred from the present study that long term follow-up studies are required to evaluate the potential of these methods and specific positive outcomes.

  9. Expansion of bovine chondrocytes on microcarriers enhances redifferentiation

    NARCIS (Netherlands)

    Malda, J.; van Blitterswijk, Clemens; Grojec, M.; Martens, D.E.; Tramper, J.; Riesle, J.U.

    2003-01-01

    Functional cartilage implants for orthopedic surgery or in vitro tissue evaluation can be created from expanded chondrocytes and biodegradable scaffolds. Expansion of chondrocytes in two-dimensional culture systems results in their dedifferentiation. The hallmark of this process is the switch of

  10. Chondrocyte behavior on nanostructured micropillar polypropylene and polystyrene surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Prittinen, Juha [Department of Applied Physics, University of Eastern Finland, Kuopio (Finland); Jiang, Yu [Department of Chemistry, University of Eastern Finland, Joensuu (Finland); Ylärinne, Janne H. [Department of Applied Physics, University of Eastern Finland, Kuopio (Finland); Pakkanen, Tapani A. [Department of Chemistry, University of Eastern Finland, Joensuu (Finland); Lammi, Mikko J., E-mail: mikko.lammi@uef.fi [Department of Applied Physics, University of Eastern Finland, Kuopio (Finland); Qu, Chengjuan [Department of Applied Physics, University of Eastern Finland, Kuopio (Finland)

    2014-10-01

    This study was aimed to investigate whether patterned polypropylene (PP) or polystyrene (PS) could enhance the chondrocytes' extracellular matrix (ECM) production and phenotype maintenance. Bovine primary chondrocytes were cultured on smooth PP and PS, as well as on nanostructured micropillar PP (patterned PP) and PS (patterned PS) for 2 weeks. Subsequently, the samples were collected for fluorescein diacetate-based cell viability tests, for immunocytochemical assays of types I and II collagen, actin and vinculin, for scanning electronic microscopic analysis of cell morphology and distribution, and for gene expression assays of Sox9, aggrecan, procollagen α{sub 1}(II), procollagen α{sub 1}(X), and procollagen α{sub 2}(I) using quantitative RT-PCR assays. After two weeks of culture, the bovine primary chondrocytes had attached on both patterned PP and PS, while practically no adhesion was observed on smooth PP. However, the best adhesion of the cells was on smooth PS. The cells, which attached on patterned PP and PS surfaces synthesized types I and II collagen. The chondrocytes' morphology was extended, and an abundant ECM network formed around the attached chondrocytes on both patterned PP and PS. Upon passaging, no significant differences on the chondrocyte-specific gene expression were observed, although the highest expression level of aggrecan was observed on the patterned PS in passage 1 chondrocytes, and the expression level of procollagen α{sub 1}(II) appeared to decrease in passaged chondrocytes. However, the expressions of procollagen α{sub 2}(I) were increased in all passaged cell cultures. In conclusion, the bovine primary chondrocytes could be grown on patterned PS and PP surfaces, and they produced extracellular matrix network around the adhered cells. However, neither the patterned PS nor PP could prevent the dedifferentiation of chondrocytes. - Highlights: • Methods to avoid chondrocyte dedifferentiation would be useful for cartilage

  11. Biomarkers of Chondrocyte Apoptosis and Autophagy in Osteoarthritis

    Science.gov (United States)

    Musumeci, Giuseppe; Castrogiovanni, Paola; Trovato, Francesca Maria; Weinberg, Annelie Martina; Al-Wasiyah, Mohammad K.; Alqahtani, Mohammed H.; Mobasheri, Ali

    2015-01-01

    Cell death with morphological and molecular features of apoptosis has been detected in osteoarthritic (OA) cartilage, which suggests a key role for chondrocyte death/survival in the pathogenesis of OA. Identification of biomarkers of chondrocyte apoptosis may facilitate the development of novel therapies that may eliminate the cause or, at least, slow down the degenerative processes in OA. The aim of this review was to explore the molecular markers and signals that induce chondrocyte apoptosis in OA. A literature search was conducted in PubMed, Scopus, Web of Science and Google Scholar using the keywords chondrocyte death, apoptosis, osteoarthritis, autophagy and biomarker. Several molecules considered to be markers of chondrocyte apoptosis will be discussed in this brief review. Molecular markers and signalling pathways associated with chondroycte apoptosis may turn out to be therapeutic targets in OA and approaches aimed at neutralizing apoptosis-inducing molecules may at least delay the progression of cartilage degeneration in OA. PMID:26334269

  12. Cause of Sudden Cardiac Deaths on Autopsy Findings; a Four-Year Report

    Directory of Open Access Journals (Sweden)

    Dinesh Rao

    2014-03-01

    Full Text Available Introduction: Incidence of sudden cardiac death (SCD has been steadily increasing all over the world. While knowing the cause of SCD is one of the favorites of the physicians involved with these cases, it is very difficult and challenging task for the forensic physician. The present report is a prospective study regarding cause of SCDs on autopsy examination in four-year period, Bangalore, India. Methods: The present prospective study is based on autopsy observations, carried out for four-year period from 2008 to 2011, and analyzed for cause of SCDs. The cases were chosen as per the definition of sudden death and autopsied. The material was divided into natural and unnatural groups. Finally, on histopathology, gross examination, hospital details, circumstantial, and police reports the cause of death was inferred. Results: A total of 2449 autopsy was conducted of which 204 cases were due to SCD. The highest SCDs were reported in 50-60 years age group (62.24%; n-127, followed closely by the age group 60-69 (28.43%; n-58. Male to female ratio was around 10:1. The maximum number of deaths (n=78 was within few hours (6 hours after the onset of signs and symptoms. In 24 (11.8% cases major narrowing was noted in both the main coronaries, in 87 (42.6% cases in the left anterior descending coronary artery (LAD, and in 18 (51.5% cases in the right coronary artery (RCA. The major cardiac pathology resulting in sudden death was coronary artery disease (n-116; 56.86% and myocardial infarction (n-104; 50.9%. most of the SCDs occurred in the place of residence (n-80; 39.2% followed closely by death in hospital (n-49; 24.01%. Conclusion: Coronary occlusion was the major contributory cause of sudden death with cardiac origin and the highest number of deaths were reported in the age 50-59 years with male to female ratio of 10:1. 

  13. MSX2 stimulates chondrocyte maturation by controlling Ihh expression.

    Science.gov (United States)

    Amano, Katsuhiko; Ichida, Fumitaka; Sugita, Atsushi; Hata, Kenji; Wada, Masahiro; Takigawa, Yoko; Nakanishi, Masako; Kogo, Mikihiko; Nishimura, Riko; Yoneda, Toshiyuki

    2008-10-24

    Several studies indicated that a homeobox gene, Msx2, is implicated in regulation of skeletal development by controlling enchondral ossification as well as membranous ossification. However, the molecular basis by which Msx2 conducts chondrogenesis is currently unclear. In this study, we examined the role of Msx2 in chondrocyte differentiation using mouse primary chondrocytes and embryonic metatarsal explants. Treatment with BMP2 up-regulated the expression of Msx2 mRNA along with chondrocyte differentiation in murine primary chondrocytes. Overexpression of wild-type Msx2 stimulated calcification of primary chondrocytes in the presence of BMP2. We also found that constitutively active Msx2 (caMsx2) enhanced BMP2-dependent calcification more efficiently than wild-type Msx2. Consistently, caMsx2 overexpression up-regulated the expression of alkaline phosphatase and collagen type X induced by BMP2. Furthermore, organ culture experiments using mouse embryonic metatarsals indicated that caMsx2 clearly stimulated the maturation of chondrocytes into the prehypertrophic and hypertrophic stages in the presence of BMP2. In contrast, knockdown of Msx2 inhibited maturation of primary chondrocytes. The stimulatory effect of Msx2 on chondrocyte maturation was enhanced by overexpression of Smad1 and Smad4 but inhibited by Smad6, an inhibitory Smad for BMP2 signaling. These data suggest that Msx2 requires BMP2/Smad signaling for its chondrogenic action. In addition, caMsx2 overexpression induced Ihh (Indian hedgehog) expression in mouse primary chondrocytes. Importantly, treatment with cyclopamine, a specific inhibitor for hedgehogs, blocked Msx2-induced chondrogenesis. Collectively, our results indicated that Msx2 promotes the maturation of chondrocytes, at least in part, through up-regulating Ihh expression.

  14. 5-Aza-2'-deoxycytidine acts as a modulator of chondrocyte hypertrophy and maturation in chick caudal region chondrocytes in culture.

    Science.gov (United States)

    Haq, Samina Hyder

    2016-06-01

    This study was carried out to explore the effect of DNA hypomethylation on chondrocytes phenotype, in particular the effect on chondrocyte hypertrophy, maturation, and apoptosis. Chondrocytes derived from caudal region of day 17 embryonic chick sterna were pretreated with hypomethylating drug 5-aza-2'-deoxycytidine for 48 hours and then maintained in the normal culture medium for up to 14 days. Histological studies showed distinct morphological changes occurred in the pretreated cultures when compared to the control cultures. The pretreated chondrocytes after 7 days in culture became bigger in size and acquired more flattened fibroblastic phenotype as well as a loss of cartilage specific extracellular matrix. Scanning electron microscopy at day 7 showed chondrocytes to have increased in cell volume and at day 14 in culture the extracellular matrix of the pretreated cultures showed regular fibrillar structure heavily embedded with matrix vesicles, which is the characteristic feature of chondrocyte hypertrophy. Transmission electron microscopic studies indicated the terminal fate of the hypertrophic cells in culture. The pretreated chondrocytes grown for 14 days in culture showed two types of cells: dark cells which had condense chromatin in dark patches and dark cytoplasm. The other light chondrocytes appeared to be heavily loaded with endoplasmic reticulum indicative of very active protein and secretory activity; their cytoplasm had large vacuoles and disintegrating cytoplasm. The biosynthetic profile showed that the pretreated cultures were actively synthesizing and secreting type X collagen and alkaline phosphatase as a major biosynthetic product.

  15. THE INFORMATION-COMMUNICATION TECHNOLOGY IN THE HOME ENVIRONMENT OF FOUR-YEAR-OLD PRESCHOOL CHILDREN

    Directory of Open Access Journals (Sweden)

    Jurka Lepičnik Vodopivec

    2014-01-01

    Full Text Available Today, preschool children live in an era of information-communication technologies (ICT. There exist various definitions of ICT. For the purposes of our research we thus used a broader definition of ICT, which stretches beyond mobiles and computers, and includes a variety of everyday technologies. Children already encounter them in their family environment. We wanted to find out which ICT children use the most, how they access them and if ICT has any influence on their development. Our sample consisted of 130 parents of four-year-old children who visit kindergarten. We were interested in the adults' – parents' influence and in the influence of the children’s gender on the access to and usage of ICT. We similarly wanted to know if these are influenced by the toy industry market and family values. A part of the research is also dedicated to the ICT’s influence on the perception of childhood and a plausible occurrence of the digital divide.

  16. Who Benefits From Four-Year Higher Education in Turkey? A Case Study of Law School

    Directory of Open Access Journals (Sweden)

    Filiz GÖLPEK

    2013-01-01

    Full Text Available In many countries, a higher education system which is based on welfare state practices prevails. In this system, the services are substantially provided by the state free of charge or at quite low prices. Since the services are financed by taxes, additional sources cannot be provided; and because of the fact that it doesn't entail a real cost benefit comparison by the users, it leads to inefficiency. From the early 80s on, there has been a tendency towards the marketing mechanisms in the provision of many public services such as education due to both strong theoretical and empirical reason, and the restrictions in public financing. Together with this tendency, the question of who benefits from education services is tried to be answered in order to discuss the stimulation of activities and additional resources on a reasonable base. The aim of this study is to determine whether it is the individual or the society who benefits from this public service at the higher education level by calculating private and social return rates through internal return rate technique which is one of the cost benefit analysis method. Findings from calculations have shown that the individual gains more than the society after a four-year university education.

  17. FOUR-YEAR-OLD NAMSAN TUNNEL CONGESTION PRICING SCHEME IN SEOUL

    Directory of Open Access Journals (Sweden)

    Bongsoo SON, Ph. D.

    2002-01-01

    Full Text Available The purpose of this paper is to evaluate the effectiveness of the congestion pricing scheme at Namsan #1 and #3 tunnels in downtown Seoul four years after its implementation. The effectiveness of the scheme was measured by the changes of various traffic impacts. The traffic volume of the two tunnels was reduced by up to 25% for the first month. After that time, the traffic volume started to increase again and then exceeded the previous volume level. However, average travel speed of the two tunnel corridors improved by up to 74%. The overall traffic volume of the four alternative routes was increased; nevertheless, their average travel speed increased as well. The number of carpool vehicles occupied by 3 or more persons including the driver during the peak periods was remarkably increased. Before the congestion fee charging, toll-charged vehicles amounted to 68.5% of the total traffic volume of the two tunnels, and then the share dropped to 29% afterwards. The empirical analysis results for the effectiveness of the congestion pricing scheme are very promising.

  18. [Four year follow-up of a screening program for prostate cancer in workers].

    Science.gov (United States)

    Reinoso-Barbero, Luis; Díaz-Garrido, Ramón; Piñaga-Solé, Montserrat; Fernández-Fernández, Miguel; Belanger-Quintana, Diego; Gómez-Gallego, Félix

    2013-01-01

    To analyze our four-year follow-up experience (2008-2011) with a prostate cancer screening program offered to employees of a banking company. Data were obtained from the health examinations carried out by the bank's in-house occupational health service (with centers in Barcelona, Madrid and Valencia). PSA (prostate-specific antigen) blood levels were measured and cases with high levels (>4 ng/ml) were followed through diagnosis and treatment, including a telephone survey of confirmed cases. Personal and occupational characteristics of the participants were analyzed as well. 750 workers (99% with administrative and/or commercial jobs) met the inclusion criteria for the screening program. Of these, 110 had elevated PSA levels on at least one occasion. The diagnosis of prostate cancer was confirmed in 21 cases. There were no associations between a diagnosis of cancer and the remaining analyzed variables. Urology and pathology records were retrieved for 76% of the contacted cases. The most frequent histological type was adenocarcinoma (98%), the most common Gleason grade at diagnosis was 6-7% (88%), and the majority of cases were treated surgically (90%).With respect to adverse effects, 48% of cases described erectile dysfunction and 33% reported urinary incontinence. In our program the observed prevalence of prostate cancer was above that expected (respectively, 21 confirmed cases vs. 12 expected). The identified cases unanimously expressed their support for the screening program. Copyright belongs to the Societat Catalana de Seguretat i Medicina del Treball.

  19. Radioactive contamination status of an elementary school in southern Miyagi Prefecture, Japan, four years after decontamination

    International Nuclear Information System (INIS)

    Yamaguchi, Toshiro; Sato, Mitsuyoshi; Nagakubo, Kazuyoshi

    2017-01-01

    We investigated the radioactive contamination status of an elementary school in southern Miyagi Prefecture, Japan, four years after decontamination. The radioactive dose rate was measured in the playground (n = 36), back yard (n = 1), parking lot (n = 3), and gutters (n = 30) with a thallium-activated sodium iodide scintillation detector. In addition, topsoil from the playground and gutters sediment were measured with a high-purity germanium semiconductor detector, and sand from the parking lot and sediment from gutters was imaged following an autoradiography method. The mean radioactive dose rate in the playground was almost 0.08 μSv/h. The dose rate was higher at sites facing the mountains, near the main gate, and by the front entrance of the school building. Meanwhile, the radioactive cesium (Cs) concentration of sediment imaged using autoradiography was > 8,000 Bq/kg. These results indicated that the radioactive dose rates in the decontaminated school were below the threshold for being a health hazard. However, the topsoil in the playground had been re-contaminated with radioactive Cs, which had likely been transported via dirt attached to children's shoes and car tires. In addition, the radioactive sediment in the gutters had likely been contaminated by rainwater, suggesting that radioactive protection is necessary when handling gutter sediment. (author)

  20. Progressive Incrementalism: U.S Foreign Economic Policy Over the Next Four Years

    Directory of Open Access Journals (Sweden)

    John M. Curtis

    2013-01-01

    Full Text Available As we look ahead following last year’s U.S. presidential and congressional elections, Canadians — and Canadian business in particular — will want to have a better sense of the economic prospects, as well as the context, of U.S. foreign economic policy over the coming half-decade or so. For, notwithstanding all the talk, debates, op-ed articles, thinktank-pieces and even books in recent years suggesting or asserting the decline of American economic power, Canada’s ever-present integration and interdependence with our southern neighbour is both a reality and now, again, an important, medium-term asset. Over the coming four years and likely beyond, and once it has completed sorting out its “fiscal cliff” matter, the U.S. is positioned to be the source of increasing economic growth. The American economy has strengths: its high productivity and innovation compared to most other countries, massive piles of private sector cash leading to all sorts of investment potential, a devalued currency, and a relatively young workforce compared to the European Union, Japan, and even Canada. Therefore, although the U.S. economy continues to face large challenges related to fiscal imbalances, there are reasons to expect increasing economic growth over the medium term.

  1. The differential prevalence of obesity and related behaviors in two- vs. four-year colleges.

    Science.gov (United States)

    N Laska, Melissa; Pasch, Keryn E; Lust, Katherine; Story, Mary; Ehlinger, Ed

    2011-02-01

    The objective of this study was to determine whether obesity prevalence and weight-related behaviors (e.g., diet, physical activity) differ among students enrolled in 2-year community/technical colleges and those attending 4-year colleges/universities. This information could inform the development of intervention strategies. Through an existing surveillance system of Minnesota postsecondary education institutions, survey data were collected from 16,539 students from 27 campuses (14 two-year college campuses, 13 four-year college/university campuses; 2007-2008), including self-reported physical activity, media use, dietary patterns, weight control behaviors, height, and weight. Unadjusted analyses indicated that students enrolled in 2-year colleges, particularly females, had a higher prevalence of overweight/obesity, lower levels of physical activity, more television viewing, higher intakes of soda, fast food, and diet pills compared to students attending 4-year colleges (P students there were fewer differences between 2-year and 4-year colleges. Controlling for sociodemographic factors (e.g., race/ethnicity, age), most disparities in prevalence estimates remained, though many were attenuated. Overall, few young adults engage in weight-related behaviors consistent with national recommendations. Two-year college students may represent a particularly at-risk group. Disparities between 2- and 4-year college students exist beyond the sociodemographic differences in these populations. Effective weight-related interventions are needed for young adults, particularly females attending 2-year colleges and all males attending postsecondary institutions.

  2. Four-year evaluation of workers exposed to trimellitic anhydride. A brief report

    International Nuclear Information System (INIS)

    McGrath, K.G.; Roach, D.; Zeiss, C.R.; Patterson, R.

    1984-01-01

    In a four-year clinical, immunologic, and environmental study of trimellitic anhydride (TMA) exposure in a single plant, 20 workers exposed to TMA powder were evaluated in 1979 and a total of 32 workers were evaluated from 1979 to 1983. Two distinct groups emerged before and after workplace control improvements were made in 1979. Seventeen of the original 20 workers were available for longitudinal study through 1983. Annual clinical evaluations and serum radioimmunoassays for total antibody binding and specific IgE binding to 125I TM-HSA (human serum albumin) were performed on all 32 workers. In 1979, six workers had antibody against TM-HSA, three had the late respiratory systemic syndrome, and two had TMA-induced allergic rhinitis or allergic rhinitis and asthma. One worker had antibody against TM-HSA without illness. Fifteen additional workers were evaluated longitudinally after institution of several workplace control measures. Four of these 15 workers had TMA exposure prior to environmental improvement and joined the study in 1982. The remaining 11 workers joined the study in 1982 and had at least two years of TMA exposure in the modified workplace. None of these 11 workers developed a TMA-induced immunologic syndrome or significant total or specific IgE antibody binding to 125I TM-HSA

  3. The Effects of Inquiry-Based Integrated Information Literacy Instruction: Four-Year Trends

    Directory of Open Access Journals (Sweden)

    Lin Ching Chen

    2014-07-01

    Full Text Available The purpose of this study was to examine the effects of four-year integrated information literacy instruction via a framework of inquiry-based learning on elementary students’ memory and comprehension. Moderating factors of students’ academic achievement was another focus of this study. The subjects were 72 students who have participated in this study since they entered an elementary school in Chiayi district. This elementary school adopted the integrated information literacy instruction, designed by the researchers and elementary school teachers, and integrated it into various subject matters via a framework of inquiry-based learning, such as Super 3 and Big6 models. A series of inquiry-based integrated information literacy instruction has been implemented since the second semester of the subjects’ first grade. A total of seven inquiry learning projects has been implemented from grade one through grade four. Fourteen instruments were used as pretests and posttests to assess students’ factual recall and conceptual understanding of subject contents in different projects. The results showed that inquiry-based integrated information literacy instruction couldhelp students memorize facts and comprehend concepts of subject contents. Regardless ofacademic achievements, if students would like to devote their efforts to inquiry processes, their memory and comprehension of subject contents improvedeffectively. However, students of low-academic achievement might need more time to be familiar with the inquiry-based learning strategy.

  4. Psychological Processes and Repeat Suicidal Behavior: A Four-Year Prospective Study

    Science.gov (United States)

    2013-01-01

    Objective: Although suicidal behavior is a major public health concern, understanding of individually sensitive suicide risk mechanisms is limited. In this study, the authors investigated, for the first time, the utility of defeat and entrapment in predicting repeat suicidal behavior in a sample of suicide attempters. Method: Seventy patients hospitalized after a suicide attempt completed a range of clinical and psychological measures (depression, hopelessness, suicidal ideation, defeat, and entrapment) while in hospital. Four years later, a nationally linked database was used to determine who had been hospitalized again after a suicide attempt. Results: Over 4 years, 24.6% of linked participants were readmitted to hospital after a suicidal attempt. In univariate logistic regression analyses, defeat and entrapment as well as depression, hopelessness, past suicide attempts, and suicidal ideation all predicted suicidal behavior over this interval. However, in the multivariate analysis, entrapment and past frequency of suicide attempts were the only significant predictors of suicidal behavior. Conclusions: This longitudinal study supports the utility of a new theoretical model in the prediction of suicidal behavior. Individually sensitive suicide risk processes like entrapment could usefully be targeted in treatment interventions to reduce the risk of repeat suicidal behavior in those who have been previously hospitalized after a suicide attempt. PMID:23855989

  5. The Impact of Impulsivity on Weight Loss Four Years after Bariatric Surgery

    Directory of Open Access Journals (Sweden)

    Kathrin Schag

    2016-11-01

    Full Text Available Bariatric surgery has serious implications on metabolic health. The reasons for a failure of bariatric surgery, i.e., limited weight loss, are multifactorial and include psychological factors. We established a theoretical model of how impulsivity is related to weight loss outcome. We propose that depressive symptoms act as a mediator between impulsivity and pathological eating behavior, and that pathological eating behavior has a direct impact on weight loss outcome. We calculated excessive weight loss (%EWL and assessed self-reported impulsivity (using the Baratt Impulsiveness Scale (BIS-15 total score, depressive symptoms (the Patient Health Questionnaire (PHQ-9 score, and pathological eating behavior (the Eating Disorder Inventory 2 (EDI-2 total score in 65 patients four years after laparoscopic sleeve gastrectomy. Regression and mediation analyses were computed to validate the theoretical model. The BIS-15, PHQ-9, and EDI-2 have medium to high correlations between each other, and EDI-2 correlated with %EWL. The mediation analysis yielded that the PHQ-9 represents a significant mediator between BIS-15 and EDI-2. The regression model between EDI-2 and %EWL was also significant. These results support our theoretical model, i.e., suggest that impulsivity has an indirect impact on weight loss outcome after bariatric surgery, mediated by depression and transferred through pathological eating behavior. Thus, the underlying psychological factors should be addressed in post-operative care to optimize weight loss outcome.

  6. Autogenous cultured growth plate chondrocyte transplantation in the treatment of physeal injury in rabbits.

    Science.gov (United States)

    Tomaszewski, R; Bohosiewicz, J; Gap, A; Bursig, H; Wysocka, A

    2014-11-01

    The aim of this experimental study on New Zealand's white rabbits was to investigate the transplantation of autogenous growth plate cells in order to treat the injured growth plate. They were assessed in terms of measurements of radiological tibial varus and histological characteristics. An experimental model of plate growth medial partial resection of the tibia in 14 New Zealand white rabbits was created. During this surgical procedure the plate growth cells were collected and cultured. While the second surgery was being performed, the autologous cultured growth plate cells were grafted at the right tibia, whereas the left tibia was used as a control group. Histological examinations showed that the grafted right tibia presented the regular shape of the plate growth with hypertrophic maturation, chondrocyte columniation and endochondral calcification. Radiological study shows that the mean tibial deformity at the left angle was 20.29° (6.25 to 33) and 7.21° (5 to 10) in the right angle. This study has demonstrated that grafting of autogenous cultured growth plate cells into a defect of the medial aspect of the proximal tibial physis can prevent bone bridge formation, growth arrest and the development of varus deformity. Cite this article: Bone Joint Res 2014;3:310-16. ©2014 The British Editorial Society of Bone & Joint Surgery.

  7. Biochar in vineyards: impact on soil quality and crop yield four years after the application

    Science.gov (United States)

    Ferreira, Carla; Verheijen, Frank; Puga, João; Keizer, Jacob; Ferreira, António

    2017-04-01

    Biochar is a recalcitrant organic carbon compound, created by biomass heating at high temperatures (300-1000°C) under low oxygen concentrations. Biochar application to agricultural soils has received increasing attention over the last years, due to its climate change mitigation and adaptation potential and reported improved soil properties and functions relevant to agronomic and environmental performance. Reported impacts are linked with increased cation exchange capacity, enhanced nutrient and water retention, and positive influences on soil microbial communities, which influence crop yields. Nevertheless, few studies have focused on mid-to-long term impacts of biochar application. This study investigated the impact of biochar on soil quality and crop yield four years after biochar application in a vineyard in North-Central Portugal. The site has a Mediterranean climate with a strong Atlantic Ocean influence, with mean annual rainfall and temperature of 1100 mm and 15°C, respectively. The soil is a relatively deep ( 80cm) sandy loam Cambisol, with gentle slopes (3°). The experimental design included three treatments: (i) control, without biochar; (ii) high biochar application rate (40 ton/ha); and (iii) biochar compost (40 ton/ha, 10% biochar). Three plots per treatment (2m×3m) were installed in March 2012, using a mini-rotavator (0-15cm depth). In May 2016, soil quality was also assessed through soil surveys and sampling. Penetration resistance was performed at the soil surface with a pocket penetrometer, and soil surface sampling rings were used for bulk density analyses (100 cm3). Bulked soil samples (0-30 cm) were collected in each plot for aggregate stability, microbial biomass (by chloroform fumigation extraction) and net mineralization rate (through photometric determination of non-incubated and incubated samples). Decomposition rate and litter stabilisation was assessed over a 3-month period through the Tea Bag Index (Keuskamp et al., 2013). The number

  8. Stomatal response of Pinus sylvestriformis to elevated CO2 concentrations during the four years of exposure

    Institute of Scientific and Technical Information of China (English)

    ZHOU Yu-mei; HAN Shi-jie; LIU Ying; JIA Xia

    2005-01-01

    Four-year-old Pinus sylvestriformis were exposed for four growing seasons in open top chambers to ambient CO2 concentration (approx. 350 μmol·mol-1) and high CO2 concentrations (500 and 700 μmol·mol-1) at Research Station of Changbai Mountain Forest Ecosystems, Chinese Academy of Sciences at Antu Town, Jilin Province, China (42oN, 128oE). Stomatal response to elevated CO2 concentrations was examined by stomatal conductance (gs), ratio of intercellular to ambient CO2 concentration (ci/ca) and stomatal number. Reciprocal transfer experiments of stomatal conductance showed that stomatal conductance in high-[CO2]-grown plants increased in comparison with ambient-[CO2]-grown plants when measured at their respective growth CO2 concentration and at the same measurement CO2 concentration (except a reduction in 700 μmol·mol-1 CO2 grown plants compared with plants on unchambered field when measured at growth CO2 concentration and 350 μmol·mol-1CO2). High-[CO2]-grown plants exhibited lower ci/ca ratios than ambient-[CO2]-grown plants when measured at their respective growth CO2 concentration. However, ci/ca ratios increased for plants grown in high CO2 concentrations compared with control plants when measured at the same CO2 concentration. There was no significant difference in stomatal number per unit long needle between elevated and ambient CO2. However, elevated CO2 concentrations reduced the total stomatal number of whole needle by the decline of stomatal line and changed the allocation pattern of stomata between upper and lower surface of needle.

  9. Injuries in Collegiate Women’s Volleyball: A Four-Year Retrospective Analysis

    Science.gov (United States)

    Sole, Christopher J.; Kavanaugh, Ashley A.; Stone, Michael H.

    2017-01-01

    A four-year retrospective analysis of injury data was conducted on a collegiate (NCAA Division I) women’s volleyball team. Twenty athletes (Year 1: age = 19.4 ± 0.9 y, height = 175.2 ± 5.1 cm, body mass = 70.5 ± 10.2 kg; Year 2: age = 20.1 ± 1.0 y, height = 175.7 ± 4.7 cm, body mass = 69.5 ± 10.1 kg; Year 3: age = 20.1 ± 1.4 y, height = 173.8 ± 6.3 cm, body mass = 69.9 ± 10.8 kg; Year 4: age = 19.5 ± 1.4 y, height = 174.4 ± 8.6 cm, body mass = 72.7 ± 10.8 kg) participated in this study, accounting for 1483 total training exposures. Injury was defined as any damage to a body part, incurred during volleyball or strength and conditioning-related activities, which interfered with training and/or competition. Injury rate was normalized to the number of athletes and exposure and expressed as injuries per 1000 exposures. A total of 133 injuries were recorded. The most common injury was to the knee (left = 7.5%, right = 12.0%). Injuries occurred most often in volleyball practice (75.2%), followed by competition (20.3%), and strength and conditioning-related activities (4.5%). Non-contact injuries (upper body = 26.3%, lower body = 53.4%) were more common than contact injuries (upper-body = 13.5%, lower-body = 6.8%). An examination of injury rates relative to the training year revealed patterns in injury occurrence. Specifically, spikes in injury rate were consistently observed during periods of increased training volume that were preceded by breaks in organized training, such as the early pre-season and off-season training periods.

  10. A four-year cardiovascular risk score for type 2 diabetic inpatients

    Directory of Open Access Journals (Sweden)

    Dolores Ramírez-Prado

    2015-06-01

    Full Text Available As cardiovascular risk tables currently in use were constructed using data from the general population, the cardiovascular risk of patients admitted via the hospital emergency department may be underestimated. Accordingly, we constructed a predictive model for the appearance of cardiovascular diseases in patients with type 2 diabetes admitted via the emergency department. We undertook a four-year follow-up of a cohort of 112 adult patients with type 2 diabetes admitted via the emergency department for any cause except patients admitted with acute myocardial infarction, stroke, cancer, or a palliative status. The sample was selected randomly between 2010 and 2012. The primary outcome was time to cardiovascular disease. Other variables (at baseline were gender, age, heart failure, renal failure, depression, asthma/chronic obstructive pulmonary disease, hypertension, dyslipidaemia, insulin, smoking, admission for cardiovascular causes, pills per day, walking habit, fasting blood glucose and creatinine. A cardiovascular risk table was constructed based on the score to estimate the likelihood of cardiovascular disease. Risk groups were established and the c-statistic was calculated. Over a mean follow-up of 2.31 years, 39 patients had cardiovascular disease (34.8%, 95% CI [26.0–43.6%]. Predictive factors were gender, age, hypertension, renal failure, insulin, admission due to cardiovascular reasons and walking habit. The c-statistic was 0.734 (standard error: 0.049. After validation, this study will provide a tool for the primary health care services to enable the short-term prediction of cardiovascular disease after hospital discharge in patients with type 2 diabetes admitted via the emergency department.

  11. Traumatic dental injuries in a university hospital: a four-year retrospective study.

    Science.gov (United States)

    Mahmoodi, Benjamin; Rahimi-Nedjat, Roman; Weusmann, Jens; Azaripour, Adriano; Walter, Christian; Willershausen, Brita

    2015-11-04

    Traumatic dental injuries present complex injuries of the dentoalveolar system. Aim of this study was to investigate the frequency and patterns of traumatic dental injuries in a University dental emergency service over four years. A retrospective investigation on all dental trauma patients presenting at the dental emergency service of the University Medical Center Mainz, Germany between 01/2010 and 12/2013 was conducted. Demographic data, the cause and type of trauma and the initial therapy were analyzed. Out of 16,301 patients, 1,305 patients (8 %; average age 14.7 years ±15.7; 60.1 % male, 39.9 % female) came due to trauma. 63.9 % of the traumas occurred on weekends. The most frequent reason for injuries was falls (54.6 %). No correlation could be found between the cause and the kind of trauma. In 48.6 % of the cases only one tooth was involved, in 33.5 % two. The permanent dentition was traumatized in 56.6 % of cases, the deciduous teeth in 41.1 %. The most frequently affected tooth was the central upper incisor (61.0 %). Hard-tissue injuries were significantly more frequent in the permanent dentition, while periodontal injuries were seen significantly more often in the deciduous dentition. Eight percent of all patients seeking help at the dental emergency service presented with trauma, meaning that dental traumatology is one of the major topics in emergencies. To improve the quality of care, further public education, expert knowledge among dental professionals and a well-structured emergency service are necessary.

  12. Four-year-old Children Align their Preferences with those of their Peers

    Directory of Open Access Journals (Sweden)

    Laura Hennefield

    2017-07-01

    Full Text Available Children express preferences for a wide range of options, such as objects, and frequently observe the preferences that others express towards these things. However, little is know about how these initial preferences develop. The present research investigated whether one particular type of social information – other children’s preferences – influences children’s own preferences. Four-year-old children observed, via video, two boys and two girls display the same preference for one of two stickers. Each child (peer expressed liking for one sticker and dislike for the other. Then children completed two rounds of the Dictator Game, a classic resource distribution task. In each round, children distributed either 10 'liked' stickers or 10 'disliked' stickers (counterbalanced between themselves and another child who was not present. If the preferences expressed by their peers influenced children’s own preferences, children should keep more of the 'liked' than 'disliked' stickers for themselves. In line with this prediction, more children kept more liked than disliked stickers, indicating their distribution patterns were influenced by their peers’ preferences. This finding suggests that children extracted informational content about the value of the stickers from their peers and used that information to guide their own preferences. Children might also have aligned their preferences with those of their peers to facilitate social bonding and group membership. This research demonstrates the strong influence of peers on children’s developing preferences, and reveals the effect of peer influence via video – a medium that young children are frequently exposed to but often struggle to learn from in other contexts.

  13. A four-year, systems-wide intervention promoting interprofessional collaboration

    Science.gov (United States)

    2012-01-01

    Background A four-year action research study was conducted across the Australian Capital Territory health system to strengthen interprofessional collaboration (IPC) though multiple intervention activities. Methods We developed 272 substantial IPC intervention activities involving 2,407 face-to-face encounters with health system personnel. Staff attitudes toward IPC were surveyed yearly using Heinemann et al's Attitudes toward Health Care Teams and Parsell and Bligh's Readiness for Interprofessional Learning scales (RIPLS). At study's end staff assessed whether project goals were achieved. Results Of the improvement projects, 76 exhibited progress, and 57 made considerable gains in IPC. Educational workshops and feedback sessions were well received and stimulated interprofessional activities. Over time staff scores on Heinemann's Quality of Interprofessional Care subscale did not change significantly and scores on the Doctor Centrality subscale increased, contrary to predictions. Scores on the RIPLS subscales of Teamwork & Collaboration and Professional Identity did not alter. On average for the assessment items 33% of staff agreed that goals had been achieved, 10% disagreed, and 57% checked neutral. There was most agreement that the study had resulted in increased sharing of knowledge between professions and improved quality of patient care, and least agreement that between-professional rivalries had lessened and communication and trust between professions improved. Conclusions Our longitudinal interventional study of IPC involving multiple activities supporting increased IPC achieved many project-specific goals. However, improvements in attitudes over time were not demonstrated and neutral assessments predominated, highlighting the difficulties faced by studies targeting change at the systems level and over extended periods. PMID:22520869

  14. McClellan Nuclear Radiation Center (MNRC) TRIGA reactor: Four years of operations

    International Nuclear Information System (INIS)

    Heidel, C.C.; Richards, W.J.

    1994-01-01

    McClellan Air Force Base, at Sacramento, California, is headquarters for the Sacramento Air Force Logistics Center (SM-ALC). McClellan Air Force Base provides extensive inspection and maintenance capabilities for the F-111, F-1 5, and other military aircraft. Criticality of the MNRC TRIGA reactor was obtained on January 20, 1990 with 63 standard TRIGA fuel elements, three fuel-followed control rods and one air-followed control rod. Presently there are 93 fuel elements in the reactor core. The reactor can be operated at 1 MW steady state power, producing pulses up to three dollars worth of reactivity addition, and can be square waved up to 1 MW. The reactor core contains a circular grid plate and a graphite reflector assembly surrounding the core. Four tangential beam ports installed in the reflector assembly provide a thermal neutron flux to four radiography bays. The reactor tank is twenty-four (24) feet deep, seven and one-half (7.5) feet in diameter, and has a protrusion in the upper portion of the reactor tank. This protrusion is scheduled for use as a neutron thermal collimator in the future. Besides the neutron radiography capabilities, the reactor contains a pneumatic rabbit system, a central thimble, an in-core irradiation facility, and three additional cutouts that provide locations for additional irradiation facilities. The central thimble can be removed along with the B-ring locations of the upper portion of the grid plate to provide an additional and larger in-core irradiation facility. A new upper grid plate has been manufactured to expand one triangular cutout so that larger experiments can be inserted directly into the reactor core. Some operational problems experienced during the first four years of operations are the timeout of the CSC and DAC watchdogs, deterioration of the heat exchanger gaskets, and loss of thermocouples in the instrumented fuel elements. (author)

  15. Four years of international counter proliferation training: The U.S. Department of Defense's experience

    International Nuclear Information System (INIS)

    Strauss, H.J.

    2001-01-01

    Full text: Over the last four years, the U.S. Department of Defense has engaged 17 countries in the former Soviet Union, Eastern/Central Europe, and the Baltic states in two counterproliferation initiatives, i.e., the DOD/FBI and the DOD/U.S. Customs Service Counterproliferation Programs. These activities are designed to train and equip border security and law enforcement personnel to prevent, deter, and investigate incidents related to weapons of mass destruction, as well as the trafficking in chemical, nuclear, and biological weapons materials and technologies. Though these programs have begun to produce tangible successes, some recipient countries have failed to demonstrate an earnest commitment to program goals. The U.S. DOD has fielded varied training courses in the region, together with associated WMD detection equipment. In spite of demands by the political leadership in many of the engaged countries, the most successful training has proven to be the more basic rather than the advanced training. Similarly, the real equipment needs prove to be for low rather than high technology. The presentation will explore the systemic, political/military, and geographic factors contributing to this result. The U.S. Department of Defense will continue to engage participating nations in these international counterproliferation programs, and will continue to respond positively to assistance requests based on recipient country needs and honest commitment. Still there remain numerous opportunities for other donor states and international agencies to make positive contributions in the counterproliferation arena. Only with increased donor state commitment - fiscal, programmatic, and personnel - together with full donor state coordination, can international proliferation and trafficking problems be effectively deterred and resolved. (author)

  16. Four years of UAS Imagery Reveals Vegetation Change Due to Permafrost Thaw

    Science.gov (United States)

    DelGreco, J. L.; Herrick, C.; Varner, R. K.; McArthur, K. J.; McCalley, C. K.; Garnello, A.; Finnell, D.; Anderson, S. M.; Crill, P. M.; Palace, M. W.

    2017-12-01

    Warming trends in sub-arctic regions have resulted in thawing of permafrost which in turn induces change in vegetation across peatlands. Collapse of palsas (i.e. permafrost plateaus) has also been correlated to increases in methane (CH4) emissions to the atmosphere. Vegetation change provides new microenvironments that promote CH4 production and emission, specifically through plant interactions and structure. By quantifying the changes in vegetation at the landscape scale, we will be able to understand the impact of thaw on CH4 emissions in these complex and climate sensitive northern ecosystems. We combine field-based measurements of vegetation composition and high resolution Unmanned Aerial Systems (UAS) imagery to characterize vegetation change in a sub-arctic mire. At Stordalen Mire (1 km x 0.5 km), Abisko, Sweden, we flew a fixed-wing UAS in July of each year between 2014 and 2017. High precision GPS ground control points were used to georeference the imagery. Seventy-five randomized square-meter plots were measured for vegetation composition and individually classified into one of five cover types, each representing a different stage of permafrost degradation. With this training data, each year of imagery was classified by cover type. The developed cover type maps were also used to estimate CH4 emissions across the mire based on average flux CH4 rates from each cover type obtained from flux chamber measurements collected at the mire. This four year comparison of vegetation cover and methane emissions has indicated a rapid response to permafrost thaw and changes in emissions. Estimation of vegetation cover types is vital in our understanding of the evolution of northern peatlands and its future role in the global carbon cycle.

  17. Pendimethalin and oxyfluorfen degradation under two irrigation conditions over four years application.

    Science.gov (United States)

    Alister, Claudio A; Gomez, Patricio A; Rojas, Sandra; Kogan, Marcelo

    2009-05-01

    A four-year field study was conducted to determine the effect of pluviometric conditions on pendimethalin and oxyfluorfen soil dynamics. Adsorption, dissipation and soil movement were studied in a sandy loam soil from 2003 to 2007. Pendimethalin and oxyfluorfen were applied every year on August at 1.33 and 0.75 kg ha(-1), respectively. Herbicide soil concentrations were determined at 0, 10, 20, 40, 90 and 340 days after application (DAA), under two pluviometric regimens, natural rainfall and irrigated (30 mm every 15 days during the first 90 DAA). More than 74% of the herbicide applied was detected at the top 2.5 cm layer for both herbicides, and none was detected at 10 cm or deeper. Pendimethalin soil half-life ranged from 10.5 to 31.5 days, and was affected mainly by the time interval between application and the first rain event. Pendimethalin soil residues at 90 DAA fluctuated from 2.5 to 13.8% of the initial amount applied, and it decreased to 2.4 and 8.6% at 340 DAA. Oxyfluorfen was more persistent than pendimethalin as indicated by its soil half-life which ranged from 34.3 to 52.3 days, affected primarily by the rain amount at the first rainfall after application. Oxyfluorfen soil residues at 90 DAA ranged from 16.7 to 34.8% and it decreased to 3.3 and 17.9% at 340 DAA. Based on half-life values, herbicide soil residues after one year, and soil depth reached by the herbicides, we conclude that both herbicides should be considered as low risk to contaminate groundwater. However, herbicide concentration at the top 2.5 cm layer should be considered in cases where runoff or soil erosion could occur, because of the potential for surface water contamination.

  18. The wind and fire disturbance in Central European mountain spruce forests: the regeneration after four years

    Directory of Open Access Journals (Sweden)

    Monika Budzáková

    2013-03-01

    Full Text Available A strong windstorm in November 2004 resulted in a huge blown-down spruce forest area in the southern part of the Tatra National Park in the Western Carpathians in Slovakia, Central Europe. The aim of this work is to study the vegetation composition of spruce forest at differently managed sites four years after this disturbance. Four study areas were selected for this purpose: (i an area where the fallen trees were extracted and new seedlings were planted; (ii an area, which was hit by a forest fire after the extraction; (iii an area where no active management was applied; (iv a reference forest unaffected by such disturbance. A total of 100 plots were selected, 25 of each area type. The result of DCA and CCA analyses consistently indicated that after this short period the non-extracted and extracted areas are currently most similar to the reference forest area, while the fire affected area differed. A one-way ANOVA comparing species cover for the different plot sizes indicated some significant differences between the extracted and non-extracted plots. The abundance of certain species commonly occurring in spruce forests, such as Dyopteris carthusiana agg., Vaccinium myrtillus and Avenella flexuosa, correlated weli with the non-extracted plots, compared to the extracted plots. Coverage of these species was lowest on burned plots. The lowest Shannon-Wiener’s diversity values were recorded in burned plots. This was most likely a consequence of mono-dominant competitive species spread, (mainly Chamerion angustifolium which profited from the altered ecological conditions following the fire. Although some differences were also registered in the Shannon-Wiener diversity index between the remaining research plots, however these were not statistically significant. The most important results of our investigations include the extensive influence of fire disturbance on vegetation. Study revealed that the wind-disturbed area is able to regenerate

  19. Stent graft placement for dysfunctional arteriovenous grafts

    Energy Technology Data Exchange (ETDEWEB)

    Jeon, Gyeong Sik [Dept. of Radiology, CHA Bundang Medical Center, College of Medicine, CHA University, Seongnam (Korea, Republic of); Shin, Byung Seok; Ohm, Joon Young; Ahn, Moon Sang [Chungnam National University Hospital, Daejeon (Korea, Republic of)

    2015-07-15

    This study aimed to evaluate the usefulness and outcomes of stent graft use in dysfunctional arteriovenous grafts. Eleven patients who underwent stent graft placement for a dysfunctional hemodialysis graft were included in this retrospective study. Expanded polytetrafluoroethylene covered stent grafts were placed at the venous anastomosis site in case of pseudoaneurysm, venous laceration, elastic recoil or residual restenosis despite the repeated angioplasty. The patency of the arteriovenous graft was evaluated using Kaplan-Meier analysis. Primary and secondary mean patency was 363 days and 741 days. Primary patency at 3, 6, and 12 months was 82%, 73%, and 32%, respectively. Secondary patency at the 3, 6, 12, 24, and 36 months was improved to 91%, 82%, 82%, 50%, and 25%, respectively. Fractures of the stent graft were observed in 2 patients, but had no effect on the patency. Stent graft placement in dysfunctional arteriovenous graft is useful and effective in prolonging graft patency.

  20. Preventing errors in administration of parenteral drugs: the results of a four-year national patient safety program.

    NARCIS (Netherlands)

    Blok, C. de; Schilp, J.; Wagner, C.

    2013-01-01

    Objectives: To evaluate the implementation of a four-year national patient safety program concerning the parenteral drug administration process in the Netherlands. Methods: Structuring the preparation and administration process of parenteral drugs reduces the number of medication errors. A

  1. Efficacy of platelet-rich fibrin matrix on viability of diced cartilage grafts in a rabbit model.

    Science.gov (United States)

    Güler, İsmail; Billur, Deniz; Aydin, Sevim; Kocatürk, Sinan

    2015-03-01

    The objective of this study was to compare the viability of cartilage grafts embedded in platelet-rich fibrin matrix (PRFM) wrapped with no material (bare diced cartilage grafts), oxidized methylcellulose (Surgicel), or acellular dermal tissue (AlloDerm). Experimental study. In this study, six New Zealand rabbits were used. Cartilage grafts including perichondrium were excised from each ear and diced into 2-mm-by 2-mm pieces. There were four comparison groups: 1) group A, diced cartilage (not wrapped with any material); 2) group B, diced cartilage wrapped with AlloDerm; 3) group C, diced cartilage grafts wrapped with Surgicel; and 4) group D, diced cartilage wrapped with PRFM. Four cartilage grafts were implanted under the skin at the back of each rabbit. All rabbits were sacrificed at the end of 10 weeks. The cartilages were stained with hematoxylin-eosin, Masson's Trichrome, and Orcein. After that, they were evaluated for the viability of chondrocytes, collagen content, fibrillar structure of matrix, and changes in peripheral tissues. When the viability of chondrocytes, the content of fiber in matrix, and changes in peripheral tissues were compared, the cartilage embedded in the PRFM group was statistically significantly higher than in the other groups (P < 0.05). We concluded that PRFM has significant advantages in ensuring the chondrocyte viability of diced cartilage grafts. It is also biocompatible, with relatively lesser inflammation and fibrosis. © 2014 The American Laryngological, Rhinological and Otological Society, Inc.

  2. Unphysiologically high magnesium concentrations support chondrocyte proliferation and redifferentiation.

    Science.gov (United States)

    Feyerabend, Frank; Witte, Frank; Kammal, Michael; Willumeit, Regine

    2006-12-01

    The effect of unphysiologically high extracellular magnesium concentrations on chondrocytes, induced by the supplementation of magnesium sulfate, was studied using a 3-phase tissue engineering model. The experiments showed that chondrocyte proliferation and redifferentiation, on the gene and protein expression level, are enhanced. A negative influence was found during chondrogenesis where an inhibition of extracellular matrix formation was observed. In addition, a direct impact on chondrocyte metabolism, elevated magnesium concentrations also affected growth factor effectiveness by consecutive influences during chondrogenesis. All observations were dosage dependent. The results of this study indicate that magnesium may be a useful tool for cartilage tissue engineering.

  3. A Measurement of Electron Neutrino Appearance in the MINOS Experiment After Four Years of Data

    Energy Technology Data Exchange (ETDEWEB)

    Cavanaugh, Steven [Harvard Univ., Cambridge, MA (United States)

    2010-05-01

    experiment. After two years of running with 3.14 x 1020 protons-on-target (POT), a limit was set at δCP = 0 for the normal (inverted) hierarchy of sin2(2θ13) < 0.29 (0.42) at 90% C.L. This study finds after four years of data with an accumulated intensity of 7 x 1020 POT that sin2(2θ13) < 0.12 (0.20) with δCP = 0 at 90% C.L. for the normal (inverted) hierarchy.

  4. [Construction of porous hydroxyapatite (HA) block loaded with cultured chondrocytes].

    Science.gov (United States)

    Yan, M; Dang, G

    1999-07-01

    To construct a kind of bone healing enhancing implant with cultured chondrocytes bound to hydroxyapatite (HA). Chondrocytes were obtained from the costicartilage of rat and were cultured on the porous HA blocks, 3 mm x 3 mm x 4 mm size, for three and seven days. Scanning electron micrograph was taken to show whether the cells grew outside and inside the pore of HA block. The cells cultured on tiny glass sheet for 2 days were used to prove where the cells come from by in situ hybridization technique with alpha1 (II) cDNA probe. Scanning electron micrographs showed that the pores of the HA surface and inside of the blocks are filled with cultured cells, especially the longer cultured block. The cells were chondrocytes confirmed by in situ hybridization. The porous HA can be used as cell cultured substrate and chondrocyte can adhere and proliferate inside the porous HA block.

  5. bFGF influences human articular chondrocyte differentiation

    DEFF Research Database (Denmark)

    Schmal, H; Zwingmann, J; Fehrenbach, M

    2007-01-01

    BACKGROUND: The possible functional role of basic fibroblast growth factor (bFGF) in regulating the mitotic and metabolic activity of primary human articular chondrocytes was investigated. METHODS: [EF1]Chondrocytes were enzymatically isolated from femoral head cartilage, and were cultured in vitro......FGF concentrations in supernatants of primary human articular chondrocytes peaked immediately after isolation and then declined. In a dose-dependent manner, bFGF enhanced cell amplification and viability. BFGF induced a decrease in the apoptotic cell population, while the number of proliferating cells remained...... by 53%, which was correlated with diminished mRNA production. Monolayer cultured chondrocytes secreted significant amounts of aggrecan that decreased over time. Secretion of this cartilage-specific marker was further reduced by the addition of bFGF. DISCUSSION: These findings highlight the potential...

  6. Msx2 Stimulates Chondrocyte Maturation by Controlling Ihh Expression*

    OpenAIRE

    Amano, Katsuhiko; Ichida, Fumitaka; Sugita, Atsushi; Hata, Kenji; Wada, Masahiro; Takigawa, Yoko; Nakanishi, Masako; Kogo, Mikihiko; Nishimura, Riko; Yoneda, Toshiyuki

    2008-01-01

    Several studies indicated that a homeobox gene, Msx2, is implicated in regulation of skeletal development by controlling enchondral ossification as well as membranous ossification. However, the molecular basis by which Msx2 conducts chondrogenesis is currently unclear. In this study, we examined the role of Msx2 in chondrocyte differentiation using mouse primary chondrocytes and embryonic metatarsal explants. Treatment with BMP2 up-regulated the expression of Msx2 mRNA...

  7. Rabbit articular cartilage defects treated by allogenic chondrocyte transplantation

    OpenAIRE

    Boopalan, P. R. J. V. C.; Sathishkumar, Solomon; Kumar, Senthil; Chittaranjan, Samuel

    2006-01-01

    Articular cartilage defects have a poor capacity for repair. Most of the current treatment options result in the formation of fibro-cartilage, which is functionally inferior to normal hyaline articular cartilage. We studied the effectiveness of allogenic chondrocyte transplantation for focal articular cartilage defects in rabbits. Chondrocytes were cultured in vitro from cartilage harvested from the knee joints of a New Zealand White rabbit. A 3 mm defect was created in the articular cartilag...

  8. Inhibition of Chondrocyte Hypertrophy of Osteoarthritis by Disruptor Peptide

    Science.gov (United States)

    2017-07-01

    with PTHR and inhibits the pathogenic beta-catenin- mediated PTHR signaling switch. In Aim 2, we will define the role of disruptor peptide in...confirmed that disruptor peptide conjugated to penetratin can enter cells. Importantly, disruptor peptide can reverse the beta-catenin- mediated PTH... mediated PTHR signaling switch in chondrocytes. Mouse primary chondrocytes express both β-catenin and PTHR. Our data showed that Pen-dis- pep

  9. Bone Graft Alternatives

    Science.gov (United States)

    ... Spine Treatment Spondylolisthesis BLOG FIND A SPECIALIST Treatments Bone Graft Alternatives Patient Education Committee Patient Education Committee ... procedure such as spinal fusion. What Types of Bone Grafts are There? Bone grafts that are transplanted ...

  10. The properties of bioengineered chondrocyte sheets for cartilage regeneration

    Directory of Open Access Journals (Sweden)

    Ota Naoshi

    2009-03-01

    Full Text Available Abstract Background Although the clinical results of autologous chondrocyte implantation for articular cartilage defects have recently improved as a result of advanced techniques based on tissue engineering procedures, problems with cell handling and scaffold imperfections remain to be solved. A new cell-sheet technique has been developed, and is potentially able to overcome these obstacles. Chondrocyte sheets applicable to cartilage regeneration can be prepared with this cell-sheet technique using temperature-responsive culture dishes. However, for clinical application, it is necessary to evaluate the characteristics of the cells in these sheets and to identify their similarities to naive cartilage. Results The expression of SOX 9, collagen type 2, 27, integrin α10, and fibronectin genes in triple-layered chondrocyte sheets was significantly increased in comparison to those in conventional monolayer culture and in a single chondrocyte sheet, implying a nature similar to ordinary cartilage. In addition, immunohistochemistry demonstrated that collagen type II, fibronectin, and integrin α10 were present in the triple-layered chondrocyte sheets. Conclusion The results of this study indicate that these chondrocyte sheets with a consistent cartilaginous phenotype and adhesive properties may lead to a new strategy for cartilage regeneration.

  11. Multi-membrane chitosan hydrogels as chondrocytic cell bioreactors.

    Science.gov (United States)

    Ladet, S G; Tahiri, K; Montembault, A S; Domard, A J; Corvol, M-T M

    2011-08-01

    We investigated the bioactivity of new chitosan-based multi-membrane hydrogel (MMH) architectures towards chondrocyte-like cells. The microstructure of the hydrogels constituting the membranes precludes any living cell penetration, whereas their lower scale architecture allows the protein diffusion. The biological behavior of chondrocytes implanted within the MMH inter-membrane spaces was studied for 45 days in culture. Chondrocytes formed cell aggregates and proliferated without loosing their chondrogenic phenotype as illustrated by collagen II and aggrecan expressions at the mRNA and protein levels. Cells produced neo-formed alcyan blue matrix proteins filling MMH interspaces. The HiF-2α/SOX9 pattern of expression suggested that the elevated chondrocytic phenotype in MMH could be related to a better hypoxic local environment than in classical culture conditions. Pro-inflammatory markers were not expressed during the period of culture. The low level of nitric oxide accumulation within the inter-membrane spaces and in the incubation medium implied that chitosan consumed nitrites produced by entrapped chondrocytes, in relation with the decrease of its molecular weight of 50%. Our data suggest that MMH structures may be considered as complex chondrocytic cell bioreactors; "active decoys of biological media", potentially promising for various biomedical applications like the inter-vertebral disk replacement. Copyright © 2011 Elsevier Ltd. All rights reserved.

  12. Antioxidant effect of bisphosphonates and simvastatin on chondrocyte lipid peroxidation

    International Nuclear Information System (INIS)

    Dombrecht, E.J.; De Tollenaere, C.B.; Aerts, K.; Cos, P.; Schuerwegh, A.J.; Bridts, C.H.; Van Offel, J.F.; Ebo, D.G.; Stevens, W.J.; De Clerck, L.S.

    2006-01-01

    The objective of this study was to evaluate the effect of bisphosphonates (BPs) and simvastatin on chondrocyte lipid peroxidation. For this purpose, a flow cytometrical method using C11-BODIPY 581/591 was developed to detect hydroperoxide-induced lipid peroxidation in chondrocytes. Tertiary butylhydroperoxide (t-BHP) induced a time and concentration dependent increase in chondrocyte lipid peroxidation. Addition of a Fe 2+ /EDTA complex to t-BHP or hydrogen peroxide (H 2 O 2 ) clearly enhanced lipid peroxidation. The lipophilic simvastatin demonstrated a small inhibition in the chondrocyte lipid peroxidation. None of three tested BPs (clodronate, pamidronate, and risedronate) had an effect on chondrocyte lipid peroxidation induced by t-BHP. However, when Fe 2+ /EDTA complex was added to t-BHP or H 2 O 2 , BPs inhibited the lipid peroxidation process varying from 25% to 58%. This study demonstrates that BPs have antioxidant properties as iron chelators, thereby inhibiting the chondrocyte lipid peroxidation. These findings add evidence to the therapeutic potential of bisphosphonates and statins in rheumatoid arthritis

  13. Endogenous versus Exogenous Growth Factor Regulation of Articular Chondrocytes

    Science.gov (United States)

    Shi, Shuiliang; Chan, Albert G.; Mercer, Scott; Eckert, George J.; Trippel, Stephen B.

    2014-01-01

    Anabolic growth factors that regulate the function of articular chondrocytes are candidates for articular cartilage repair. Such factors may be delivered by pharmacotherapy in the form of exogenous proteins, or by gene therapy as endogenous proteins. It is unknown whether delivery method influences growth factor effectiveness in regulating articular chondrocyte reparative functions. We treated adult bovine articular chondrocytes with exogenous recombinant insulin-like growth factor-I (IGF-I) and transforming growth factor-beta1 (TGF-β1), or with the genes encoding these growth factors for endogenous production. Treatment effects were measured as change in chondrocyte DNA content, glycosaminoglycan production, and aggrecan gene expression. We found that IGF-I stimulated chondrocyte biosynthesis similarly when delivered by either exogenous or endogenous means. In contrast, exogenous TGF-ß1 stimulated these reparative functions, while endogenous TGF-ß1 had little effect. Endogenous TGF-ß1 became more bioactive following activation of the transgene protein product. These data indicate that effective mechanisms of growth factor delivery for articular cartilage repair may differ for different growth factors. In the case of IGF-I, gene therapy or protein therapy appear to be viable options. In contrast, TGF-ß1 gene therapy may be constrained by a limited ability of chondrocytes to convert latent complexes to an active form. PMID:24105960

  14. Sphingosine-1-phosphate stimulates rat primary chondrocyte proliferation

    International Nuclear Information System (INIS)

    Kim, Mi-Kyoung; Lee, Ha Young; Kwak, Jong-Young; Park, Joo-In; Yun, Jeanho; Bae, Yoe-Sik

    2006-01-01

    Rat primary chondrocytes express the sphingosine-1-phosphate (S1P) receptor, S1P 2 , S1P 3 , S1P 4 , but not S1P 1 . When chondrocytes were stimulated with S1P or phytosphingosine-1-phosphate (PhS1P, an S1P 1 - and S1P 4 -selective agonist), phospholipase C-mediated cytosolic calcium increase was dramatically induced. S1P and PhS1P also stimulated two kinds of mitogen-activated protein kinases, extracellular signal-regulated kinase (ERK) and p38 kinase in chondrocytes. In terms of the two phospholipids-mediated functional modulation of chondrocytes, S1P and PhS1P stimulated cellular proliferation. The two phospholipids-induced chondrocyte proliferations were almost completely blocked by PD98059 but not by SB203580, suggesting that ERK but not p38 kinase is essentially required for the proliferation. Pertussis toxin almost completely inhibited the two phospholipids-induced cellular proliferation and ERK activation, indicating the crucial role of G i protein. This study demonstrates the physiological role of two important phospholipids (S1P and PhS1P) on the modulation of rat primary chondrocyte proliferation, and the crucial role played by ERK in the process

  15. Endogenous versus exogenous growth factor regulation of articular chondrocytes.

    Science.gov (United States)

    Shi, Shuiliang; Chan, Albert G; Mercer, Scott; Eckert, George J; Trippel, Stephen B

    2014-01-01

    Anabolic growth factors that regulate the function of articular chondrocytes are candidates for articular cartilage repair. Such factors may be delivered by pharmacotherapy in the form of exogenous proteins, or by gene therapy as endogenous proteins. It is unknown whether delivery method influences growth factor effectiveness in regulating articular chondrocyte reparative functions. We treated adult bovine articular chondrocytes with exogenous recombinant insulin-like growth factor-I (IGF-I) and transforming growth factor-beta1 (TGF-β1), or with the genes encoding these growth factors for endogenous production. Treatment effects were measured as change in chondrocyte DNA content, glycosaminoglycan production, and aggrecan gene expression. We found that IGF-I stimulated chondrocyte biosynthesis similarly when delivered by either exogenous or endogenous means. In contrast, exogenous TGF-β1 stimulated these reparative functions, while endogenous TGF-β1 had little effect. Endogenous TGF-β1 became more bioactive following activation of the transgene protein product. These data indicate that effective mechanisms of growth factor delivery for articular cartilage repair may differ for different growth factors. In the case of IGF-I, gene therapy or protein therapy appear to be viable options. In contrast, TGF-β1 gene therapy may be constrained by a limited ability of chondrocytes to convert latent complexes to an active form. Published 2013 by Wiley Periodicals, Inc. on behalf of the Orthopaedic Research Society. This article is a U.S. Government work and is in the public domain in the USA.

  16. Weakness and numbness of extremities with bowel and bladder dysfunction for four years

    Directory of Open Access Journals (Sweden)

    Han-hui FU

    2017-03-01

    Full Text Available The patient was a 53-year-old female who was admitted for “numbness and weakness of lower extremities, urinary and defecation dysfunction for 4 years” in September 2016.  Four years ago, the patient felt numbness and weakness of lower extremities, with chest zonesthesia and back pain. The weakness of left lower extremity got severer next morning and she cannot feel her left foot on the pedal when riding bicycle. At night, she presented numbness under umbilicus level, feet scuff and urine retention. She was also unable to squat. No fever, headache, vomiting or visual defect was found. She was treated as stroke which showed no effect. Two days later, the numbness extended to thoracic level with no sweating below, dyspnea, deep voice, bucking, dysphagia and weakness of both upper and lower extremities. Spinal MRI showed a long T1 signal and long T2 signal extended cervical cord lesion from C4-T5 segments and spinal cord swelling. She was suspected to have multiple sclerosis (MS or neuromyelitis optica (NMO, thus prednisone was given that resulted in the improvement other than numbness of lower extremities, stagger and bladder dysfunction. In the following 4 years, alike symptoms relapsed every year. Though the glucocorticoid treatment was helpful for symptoms remission, sequelae including extremities numbness, weakness, stagger, urinary incontinence and intermittent constipation were left. The predisposing and relieving factors were unclear for relapse. Six months ago, the patient stopped the glucocorticoid treatment by herself, followed by the increase of attack frequencies. One month ago, she got diplopia in the right eye. The patient was admitted for further diagnosis and treatment. Since the onset of symptoms, the patient has been conscious. The appetite and defecation have been normal. No dryness of mouth and eye, shin rash, Reynolds Syndrome, photosensitization and joint pain was noticed. She has a weight loss of 5kg, during which time

  17. Snorc is a novel cartilage specific small membrane proteoglycan expressed in differentiating and articular chondrocytes

    DEFF Research Database (Denmark)

    Heinonen, J; Taipaleenmäki, H; Roering, P

    2011-01-01

    OBJECTIVE: Maintenance of chondrocyte phenotype is a major issue in prevention of degeneration and repair of articular cartilage. Although the critical pathways in chondrocyte maturation and homeostasis have been revealed, the in-depth understanding is deficient and novel modifying components...... subgroups. Cartilage specific expression was highest in proliferating and prehypertrophic zones during development, and in adult articular cartilage, expression was restricted to the uncalcified zone, including chondrocyte clusters in human osteoarthritic cartilage. Studies with experimental chondrogenesis...... chondrocytes and adult articular chondrocytes with possible functions associated with development and maintenance of chondrocyte phenotype....

  18. Chondrocytic Atf4 regulates osteoblast differentiation and function via Ihh.

    Science.gov (United States)

    Wang, Weiguang; Lian, Na; Ma, Yun; Li, Lingzhen; Gallant, Richard C; Elefteriou, Florent; Yang, Xiangli

    2012-02-01

    Atf4 is a leucine zipper-containing transcription factor that activates osteocalcin (Ocn) in osteoblasts and indian hedgehog (Ihh) in chondrocytes. The relative contribution of Atf4 in chondrocytes and osteoblasts to the regulation of skeletal development and bone formation is poorly understood. Investigations of the Atf4(-/-);Col2a1-Atf4 mouse model, in which Atf4 is selectively overexpressed in chondrocytes in an Atf4-null background, demonstrate that chondrocyte-derived Atf4 regulates osteogenesis during development and bone remodeling postnatally. Atf4 overexpression in chondrocytes of the Atf4(-/-);Col2a1-Atf4 double mutants corrects the reduction in stature and limb in Atf4(-/-) embryos and rectifies the decrease in Ihh expression, Hh signaling, proliferation and accelerated hypertrophy that characterize the Atf4(-/-) developing growth plate cartilages. Unexpectedly, this genetic manipulation also restores the expression of osteoblastic marker genes, namely Ocn and bone sialoprotein, in Atf4(-/-) developing bones. In Atf4(-/-);Col2a1-Atf4 adult mice, all the defective bone parameters found in Atf4(-/-) mice, including bone volume, trabecular number and thickness, and bone formation rate, are rescued. In addition, the conditioned media of ex vivo cultures from wild-type or Atf4(-/-);Col2a1-Atf4, but not Atf4(-/-) cartilage, corrects the differentiation defects of Atf4(-/-) bone marrow stromal cells and Ihh-blocking antibody eliminates this effect. Together, these data indicate that Atf4 in chondrocytes is required for normal Ihh expression and for its paracrine effect on osteoblast differentiation. Therefore, the cell-autonomous role of Atf4 in chondrocytes dominates the role of Atf4 in osteoblasts during development for the control of early osteogenesis and skeletal growth.

  19. The chondrocytic journey in endochondral bone growth and skeletal dysplasia.

    Science.gov (United States)

    Yeung Tsang, Kwok; Wa Tsang, Shun; Chan, Danny; Cheah, Kathryn S E

    2014-03-01

    The endochondral bones of the skeleton develop from a cartilage template and grow via a process involving a cascade of chondrocyte differentiation steps culminating in formation of a growth plate and the replacement of cartilage by bone. This process of endochondral ossification, driven by the generation of chondrocytes and their subsequent proliferation, differentiation, and production of extracellular matrix constitute a journey, deviation from which inevitably disrupts bone growth and development, and is the basis of human skeletal dysplasias with a wide range of phenotypic severity, from perinatal lethality to progressively deforming. This highly coordinated journey of chondrocyte specification and fate determination is controlled by a myriad of intrinsic and extrinsic factors. SOX9 is the master transcription factor that, in concert with varying partners along the way, directs the different phases of the journey from mesenchymal condensation, chondrogenesis, differentiation, proliferation, and maturation. Extracellular signals, including bone morphogenetic proteins, wingless-related MMTV integration site (WNT), fibroblast growth factor, Indian hedgehog, and parathyroid hormone-related peptide, are all indispensable for growth plate chondrocytes to align and organize into the appropriate columnar architecture and controls their maturation and transition to hypertrophy. Chondrocyte hypertrophy, marked by dramatic volume increase in phases, is controlled by transcription factors SOX9, Runt-related transcription factor, and FOXA2. Hypertrophic chondrocytes mediate the cartilage to bone transition and concomitantly face a live-or-die situation, a subject of much debate. We review recent insights into the coordination of the phases of the chondrocyte journey, and highlight the need for a systems level understanding of the regulatory networks that will facilitate the development of therapeutic approaches for skeletal dysplasia. Copyright © 2014 Wiley Periodicals

  20. Formation of Hyaline Cartilage Tissue by Passaged Human Osteoarthritic Chondrocytes.

    Science.gov (United States)

    Bianchi, Vanessa J; Weber, Joanna F; Waldman, Stephen D; Backstein, David; Kandel, Rita A

    2017-02-01

    When serially passaged in standard monolayer culture to expand cell number, articular chondrocytes lose their phenotype. This results in the formation of fibrocartilage when they are used clinically, thus limiting their use for cartilage repair therapies. Identifying a way to redifferentiate these cells in vitro is critical if they are to be used successfully. Transforming growth factor beta (TGFβ) family members are known to be crucial for regulating differentiation of fetal limb mesenchymal cells and mesenchymal stromal cells to chondrocytes. As passaged chondrocytes acquire a progenitor-like phenotype, the hypothesis of this study was that TGFβ supplementation will stimulate chondrocyte redifferentiation in vitro in serum-free three-dimensional (3D) culture. Human articular chondrocytes were serially passaged twice (P2) in monolayer culture. P2 cells were then placed in high-density (3D) culture on top of membranes (Millipore) and cultured for up to 6 weeks in chemically defined serum-free redifferentiation media (SFRM) in the presence or absence of TGFβ. The tissues were evaluated histologically, biochemically, by immunohistochemical staining, and biomechanically. Passaged human chondrocytes cultured in SFRM supplemented with 10 ng/mL TGFβ3 consistently formed a continuous layer of articular-like cartilage tissue rich in collagen type 2 and aggrecan and lacking collagen type 1 and X in the absence of a scaffold. The tissue developed a superficial zone characterized by expression of lubricin and clusterin with horizontally aligned collagen fibers. This study suggests that passaged human chondrocytes can be used to bioengineer a continuous layer of articular cartilage-like tissue in vitro scaffold free. Further study is required to evaluate their ability to repair cartilage defects in vivo.

  1. A Survey of Marketing and Market Research Activities in Two and Four-Year Colleges and Universities.

    Science.gov (United States)

    Smith, Janet D.; And Others

    A survey of marketing and market research activities in large, urban, two- and four-year colleges was conducted to identify the nature of marketing problems faced by institutions; current and preferred emphasis placed on market research activities; modes of assessing the needs of target groups and developing appropriate programs; the location of…

  2. A comparison of four years of limnology in the Sawtooth Lakes with emphasis on fertilization in Redfish Lake

    International Nuclear Information System (INIS)

    Budy, P.; Luecke, C.; Wurtsbaugh, W.A.

    1996-01-01

    Included in this section of the report on limnology of Lakes in the Snake River Plain are descriptions of four years of limnological sampling to compare inter and intra annual variability in lake productivity to evaluate potential rearing conditions for juvenile sockeyed salmon. Data was used to evaluate the effects of nutrient enhancement, annual weather patterns, and planktivore consumption on lake productivity

  3. Surviving and Thriving: The Adaptive Responses of U.S. Four-Year Colleges and Universities during the Great Recession

    Science.gov (United States)

    Brint, Steven; Yoshikawa, Sarah R. K.; Rotondi, Matthew B.; Viggiano, Tiffany; Maldonado, John

    2016-01-01

    Press reports and industry statistics both give incomplete pictures of the outcomes of the Great Recession for U.S. four-year colleges and universities. To address these gaps, we conducted a statistical analysis of all articles that appeared in Lexis-Nexis on a sample of more than 300 U.S. colleges and universities during the Recession years. We…

  4. Three-to Four-Year-Olds' Recognition That Symbols Have a Stable Meaning: Pictures Are Understood Before Written Words

    Science.gov (United States)

    Apperly, Ian. A.; Williams, Emily; Williams, Joelle

    2004-01-01

    In 4 experiments 120 three-to four-year-old non readers were asked the identity of a symbolic representation as it appeared with different objects. Consistent with Bialystok (2000), many children judged the identity of written words to vary according to the object with which they appeared but few made such errors with recognizable pictures.…

  5. A Phenomenological Study: The Lived Experience of Former Foster Youth Attending a Four-Year College in Southern California

    Science.gov (United States)

    Lee, Dora Yiu Lam

    2010-01-01

    This qualitative study examined the lived experience of eight individuals attending a four-year college who were all part of a campus support program for former foster youth. The purpose of this phenomenological study was to understand and explore the lived experiences of these unique college students that have gone through the foster care system.…

  6. A comparison of four years of limnology in the Sawtooth Lakes with emphasis on fertilization in Redfish Lake

    Energy Technology Data Exchange (ETDEWEB)

    Budy, P.; Luecke, C.; Wurtsbaugh, W.A.

    1996-05-01

    Included in this section of the report on limnology of Lakes in the Snake River Plain are descriptions of four years of limnological sampling to compare inter and intra annual variability in lake productivity to evaluate potential rearing conditions for juvenile sockeyed salmon. Data was used to evaluate the effects of nutrient enhancement, annual weather patterns, and planktivore consumption on lake productivity.

  7. The Role of Addressee Backchannels and Conversational Grounding in Vicarious Word Learning in Four-Year-Olds

    Science.gov (United States)

    Tolins, Jackson; Namiranian, Neda; Akhtar, Nameera; Fox Tree, Jean E.

    2017-01-01

    Children successfully learn words through overhearing others engaged in verbal interactions. The current studies investigated the degree to which four-year-old overhearers are influenced by the response behaviors of addressees and by the interactional pattern of the speakers and addressees. It was found that while addressee responses on their own…

  8. Two-, Three-, and Four-Year Follow-Up on the Self-Regulatory Treatment of Chronic Headache.

    Science.gov (United States)

    Blanchard, Edward B.; And Others

    1987-01-01

    Chronic tension and vascular headache patients, initially treated with relaxation and biofeedback, were followed-up on an annual basis at two-, three-, and four-years posttreatment. Tension headache patients generally showed good maintenance of initial headache reduction at Year Four. Vascular patients showed a nonsignificant trend for gradual…

  9. Experiences of Female Faculty with Maternity Leave at Four-Year Universities in an Upper Midwest State

    Science.gov (United States)

    Myerchin, Audra Dawn

    2012-01-01

    The purpose of this mixed methods study was to explore female faculty members' experiences with maternity leave, while working as a career academic in higher education. Participants consisted of women currently employed in an upper Midwest state at six four-year institutions. An online survey was completed by 121 women, and 30 of these women also…

  10. Transitive Relational Mappings in Three- and Four-Year-Olds: The Analogy of Goldilocks and the Three Bears.

    Science.gov (United States)

    Goswami, Usha

    1995-01-01

    In three experiments, three- and four-year olds were asked to map relative size from one array of objects to another, map relative size to relative proportion, and map relative size to a variety of perceptual dimensions. Children were able to make relational mappings based on size when spatial positions and concrete representations of size of…

  11. Four-Year Cross-Lagged Associations between Physical and Mental Health in the Medical Outcomes Study.

    Science.gov (United States)

    Hays, Ron D.; And Others

    1994-01-01

    Applied structural equation modeling to evaluation of cross-lagged panel models. Self-reports of physical and mental health at three time points spanning four-year interval were analyzed to illustrate cross-lagged analysis methodology. Data were analyzed from 856 patients with hypertension, diabetes, heart disease, or depression. Cross-lagged…

  12. Mechanical properties and structure-function relationships of human chondrocyte-seeded cartilage constructs after in vitro culture.

    Science.gov (United States)

    Middendorf, Jill M; Griffin, Darvin J; Shortkroff, Sonya; Dugopolski, Caroline; Kennedy, Stephen; Siemiatkoski, Joseph; Cohen, Itai; Bonassar, Lawrence J

    2017-10-01

    Autologous Chondrocyte Implantation (ACI) is a widely recognized method for the repair of focal cartilage defects. Despite the accepted use, problems with this technique still exist, including graft hypertrophy, damage to surrounding tissue by sutures, uneven cell distribution, and delamination. Modified ACI techniques overcome these challenges by seeding autologous chondrocytes onto a 3D scaffold and securing the graft into the defect. Many studies on these tissue engineered grafts have identified the compressive properties, but few have examined frictional and shear properties as suggested by FDA guidance. This study is the first to perform three mechanical tests (compressive, frictional, and shear) on human tissue engineered cartilage. The objective was to understand the complex mechanical behavior, function, and changes that occur with time in these constructs grown in vitro using compression, friction, and shear tests. Safranin-O histology and a DMMB assay both revealed increased sulfated glycosaminoglycan (sGAG) content in the scaffolds with increased maturity. Similarly, immunohistochemistry revealed increased lubricin localization on the construct surface. Confined compression and friction tests both revealed improved properties with increased construct maturity. Compressive properties correlated with the sGAG content, while improved friction coefficients were attributed to increased lubricin localization on the construct surfaces. In contrast, shear properties did not improve with increased culture time. This study suggests the various mechanical and biological properties of tissue engineered cartilage improve at different rates, indicating thorough mechanical evaluation of tissue engineered cartilage is critical to understanding the performance of repaired cartilage. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:2298-2306, 2017. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  13. Characterization of collagenase-3 binding and internalization by rabbit chondrocytes

    International Nuclear Information System (INIS)

    Raggatt, L.J.; Choundhury, I.; Williams, S.

    2002-01-01

    Full text: Collagenase-3 (MMP-13) is an extracellular matrix metalloproteinase that cleaves type II collagen, the major protein component of cartilage, with high specificity. Several studies have identified increased levels of MMP-13 in arthritic synovial fluid where it may contribute to matrix destruction in this disease. Our laboratory has previously documented a process where by osteoblastic cells remove MMP-13 from the surrounding milieu by binding the enzyme to a specific receptor. The enzyme is then internalized and degraded through the actions of the endocytotic receptor, the low-density lipoprotein receptor-related protein (LRP). Such a mechanism provides for a controlled elimination of a potentially destructive enzyme from the extracellular environment. This process of MMP-13 internalization also occurs in chondrocytes and is significantly reduced in OA chondrocytes. We are currently characterizing the internalization of MMP-13 in normal rabbit chondrocytes. Primary rabbit chondrocytes were harvested and cultured in monolayers for three passages. Reverse transcription polymerase chain reaction (RT-PCR) was used to asses the cell phenotype during the culture period and the rabbit chondrocytes were found to express the cartilage specific genes aggrecan and type II collagen throughout this time. 125I-MMP-13 was used to assess the ability of the rabbit chondrocytes to bind MMP-13. Appreciable specific cell-association of MMP-13 was detected after 10 mm of exposure to the ligand and equilibrium was obtained after 2 h. After identifying the time to equilibrium we determined whether binding was saturable by incubating the chondrocytes with increasing concentrations of 125I-MMP-13 ranging from 0 to 100 nM at 4 deg C for 2h. The amount of specifically associated MMP-13 approached saturation at 75 nM, allowing assessment of the receptor kinetics. Finally, we have assessed the ability of rabbit chondrocytes to internalize a single cohort of 125I-MMP-13 over time at

  14. Low-intensity pulsed ultrasound affects human articular chondrocytes in vitro

    NARCIS (Netherlands)

    Korstjens, C.M.; van der Rijt, R.H.H.; Albers, G.H.; Semeins, C.M.; Klein-Nulend, J.

    2008-01-01

    We investigated whether low-intensity pulsed ultrasound (LIPUS) stimulates chondrocyte proliferation and matrix production in explants of human articular cartilage obtained from donors suffering from unicompartimental osteoarthritis of the knee, as well as in isolated human chondrocytes in vitro.

  15. Pronounced biomaterial dependency in cartilage regeneration using nonexpanded compared with expanded chondrocytes

    NARCIS (Netherlands)

    Tsuchida, A.I.; Bekkers, J.E.J.; Beekhuizen, M.; Vonk, L.A.; Dhert, W.J.A.; Saris, Daniël B.F.; Creemers, L.B.

    2013-01-01

    We aimed to investigate freshly isolated compared with culture-expanded chondrocytes with respect to early regenerative response, cytokine production and cartilage formation in response to four commonly used biomaterials. Materials & methods: Chondrocytes were both directly and after expansion to

  16. TGF-β2 is involved in the preservation of the chondrocyte phenotype under hypoxic conditions

    NARCIS (Netherlands)

    Das, R.; Timur, U. T.; Edip, S.; Haak, E.; Wruck, C.; Weinans, H.; Jahr, H.

    2015-01-01

    Culturing chondrocytes under oxygen tension closely resembling their in vivo environment has been shown to have positive effects on matrix synthesis. In redifferentiation of expanded chondrocytes, hypoxia increased collagen type II expression. However, the mechanism by which hypoxia enhances

  17. The Signaling Pathways Involved in Chondrocyte Differentiation and Hypertrophic Differentiation

    Directory of Open Access Journals (Sweden)

    Jianmei Li

    2016-01-01

    Full Text Available Chondrocytes communicate with each other mainly via diffusible signals rather than direct cell-to-cell contact. The chondrogenic differentiation of mesenchymal stem cells (MSCs is well regulated by the interactions of varieties of growth factors, cytokines, and signaling molecules. A number of critical signaling molecules have been identified to regulate the differentiation of chondrocyte from mesenchymal progenitor cells to their terminal maturation of hypertrophic chondrocytes, including bone morphogenetic proteins (BMPs, SRY-related high-mobility group-box gene 9 (Sox9, parathyroid hormone-related peptide (PTHrP, Indian hedgehog (Ihh, fibroblast growth factor receptor 3 (FGFR3, and β-catenin. Except for these molecules, other factors such as adenosine, O2 tension, and reactive oxygen species (ROS also have a vital role in cartilage formation and chondrocyte maturation. Here, we outlined the complex transcriptional network and the function of key factors in this network that determine and regulate the genetic program of chondrogenesis and chondrocyte differentiation.

  18. Roles of Chondrocytes in Endochondral Bone Formation and Fracture Repair

    Science.gov (United States)

    Hinton, R.J.; Jing, Y.; Jing, J.; Feng, J.Q.

    2016-01-01

    The formation of the mandibular condylar cartilage (MCC) and its subchondral bone is an important but understudied topic in dental research. The current concept regarding endochondral bone formation postulates that most hypertrophic chondrocytes undergo programmed cell death prior to bone formation. Under this paradigm, the MCC and its underlying bone are thought to result from 2 closely linked but separate processes: chondrogenesis and osteogenesis. However, recent investigations using cell lineage tracing techniques have demonstrated that many, perhaps the majority, of bone cells are derived via direct transformation from chondrocytes. In this review, the authors will briefly discuss the history of this idea and describe recent studies that clearly demonstrate that the direct transformation of chondrocytes into bone cells is common in both long bone and mandibular condyle development and during bone fracture repair. The authors will also provide new evidence of a distinct difference in ossification orientation in the condylar ramus (1 ossification center) versus long bone ossification formation (2 ossification centers). Based on our recent findings and those of other laboratories, we propose a new model that contrasts the mode of bone formation in much of the mandibular ramus (chondrocyte-derived) with intramembranous bone formation of the mandibular body (non-chondrocyte-derived). PMID:27664203

  19. Articular chondrocyte alignment in the rat after surgically induced osteoarthritis

    Science.gov (United States)

    Takahashi, Hideaki; Tamaki, Hiroyuki; Yamamoto, Noriaki; Onishi, Hideaki

    2017-01-01

    [Purpose] Chondrocytes in articular cartilage are aligned as columns from the joint surface. Notably, loss of chondrocyte and abnormalities of differentiation factors give rise to osteoarthritis (OA). However, the relationship between chondrocyte alignment and OA progression remains unclear. This study was performed to investigate temporal alterations in surgically-induced OA rats. [Subjects and Methods] Thirteen-week-old Wistar rats (n=30) underwent destabilized medial meniscus surgery in their right knee and sham surgery in their left knee. Specimens (n=5) were collected at 0, 1, 2, 4 and 8 weeks after surgery. Histological analysis with Osteoarthritis Research Society International (OARSI) scores, cell density ratios, cell alignments and correlation between OARSI scores and cell density/alignment was performed. [Results] OARSI scores were significantly higher at 1, 2, 4 and 8 weeks in the DMM group than in the control. Cell density ratios were decreased significantly in the DMM group at 2, 4 and 8 weeks compared with the control. Chondrocyte alignment was decreased significantly in the DMM group at 4 and 8 weeks. There were negative correlations between OA severity and cell density / cell alignment. [Conclusion] The results suggest a relationship between chondrocyte alignment and cartilage homeostasis, which plays an important role in OA progression. PMID:28533592

  20. Autologous chondrocyte implantation: superior biologic properties of hyaline cartilage repairs.

    Science.gov (United States)

    Henderson, Ian; Lavigne, Patrick; Valenzuela, Herminio; Oakes, Barry

    2007-02-01

    Information regarding the quality of autologous chondrocyte implantation repair is needed to determine whether the current autologous chondrocyte implantation surgical technology and the subsequent biologic repair processes are capable of reliably forming durable hyaline or hyaline-like cartilage in vivo. We report and analyze the properties and qualities of autologous chondrocyte implantation repairs. We evaluated 66 autologous chondrocyte implantation repairs in 57 patients, 55 of whom had histology, indentometry, and International Cartilage Repair Society repair scoring at reoperation for mechanical symptoms or pain. International Knee Documentation Committee scores were used to address clinical outcome. Maximum stiffness, normalized stiffness, and International Cartilage Repair Society repair scoring were higher for hyaline articular cartilage repairs compared with fibrocartilage, with no difference in clinical outcome. Reoperations revealed 32 macroscopically abnormal repairs (Group B) and 23 knees with normal-looking repairs in which symptoms leading to arthroscopy were accounted for by other joint disorders (Group A). In Group A, 65% of repairs were either hyaline or hyaline-like cartilage compared with 28% in Group B. Autologous chondrocyte repairs composed of fibrocartilage showed more morphologic abnormalities and became symptomatic earlier than hyaline or hyaline-like cartilage repairs. The hyaline articular cartilage repairs had biomechanical properties comparable to surrounding cartilage and superior to those associated with fibrocartilage repairs.

  1. Bone grafting: An overview

    Directory of Open Access Journals (Sweden)

    D. O. Joshi

    2010-08-01

    Full Text Available Bone grafting is the process by which bone is transferred from a source (donor to site (recipient. Due to trauma from accidents by speedy vehicles, falling down from height or gunshot injury particularly in human being, acquired or developmental diseases like rickets, congenital defects like abnormal bone development, wearing out because of age and overuse; lead to bone loss and to replace the loss we need the bone grafting. Osteogenesis, osteoinduction, osteoconduction, mechanical supports are the four basic mechanisms of bone graft. Bone graft can be harvested from the iliac crest, proximal tibia, proximal humerus, proximal femur, ribs and sternum. An ideal bone graft material is biologically inert, source of osteogenic, act as a mechanical support, readily available, easily adaptable in terms of size, shape, length and replaced by the host bone. Except blood, bone is grafted with greater frequency. Bone graft indicated for variety of orthopedic abnormalities, comminuted fractures, delayed unions, non-unions, arthrodesis and osteomyelitis. Bone graft can be harvested from the iliac crest, proximal tibia, proximal humerus, proximal femur, ribs and sternum. By adopting different procedure of graft preservation its antigenicity can be minimized. The concept of bone banking for obtaining bone grafts and implants is very useful for clinical application. Absolute stability require for successful incorporation. Ideal bone graft must possess osteogenic, osteoinductive and osteocon-ductive properties. Cancellous bone graft is superior to cortical bone graft. Usually autologous cancellous bone graft are used as fresh grafts where as allografts are employed as an alloimplant. None of the available type of bone grafts possesses all these properties therefore, a single type of graft cannot be recomm-ended for all types of orthopedic abnormalities. Bone grafts and implants can be selected as per clinical problems, the equipments available and preference of

  2. THE EFFECT OF PIROXICAM ON THE METABOLISM OF ISOLATED HUMAN CHONDROCYTES

    NARCIS (Netherlands)

    BULSTRA, SK; KUIJER, R; BUURMAN, WA; TERWINDTROUWENHORST, E; GUELEN, PJM; VANDERLINDEN, AJ

    The effect of piroxicam on the metabolism of healthy and osteoarthrotic (OA) chondrocytes was studied in vitro. The chondrocytes were obtained from five healthy, five moderately OA, and four severely OA hips or knees. The chondrocytes were cultured in a high-density, short-term in vitro model. In

  3. Deciphering chondrocyte behaviour in matrix-induced autologous chondrocyte implantation to undergo accurate cartilage repair with hyaline matrix.

    Science.gov (United States)

    Demoor, M; Maneix, L; Ollitrault, D; Legendre, F; Duval, E; Claus, S; Mallein-Gerin, F; Moslemi, S; Boumediene, K; Galera, P

    2012-06-01

    Since the emergence in the 1990s of the autologous chondrocytes transplantation (ACT) in the treatment of cartilage defects, the technique, corresponding initially to implantation of chondrocytes, previously isolated and amplified in vitro, under a periosteal membrane, has greatly evolved. Indeed, the first generations of ACT showed their limits, with in particular the dedifferentiation of chondrocytes during the monolayer culture, inducing the synthesis of fibroblastic collagens, notably type I collagen to the detriment of type II collagen. Beyond the clinical aspect with its encouraging results, new biological substitutes must be tested to obtain a hyaline neocartilage. Therefore, the use of differentiated chondrocytes phenotypically stabilized is essential for the success of ACT at medium and long-term. That is why researchers try now to develop more reliable culture techniques, using among others, new types of biomaterials and molecules known for their chondrogenic activity, giving rise to the 4th generation of ACT. Other sources of cells, being able to follow chondrogenesis program, are also studied. The success of the cartilage regenerative medicine is based on the phenotypic status of the chondrocyte and on one of its essential component of the cartilage, type II collagen, the expression of which should be supported without induction of type I collagen. The knowledge accumulated by the scientific community and the experience of the clinicians will certainly allow to relief this technological challenge, which influence besides, the validation of such biological substitutes by the sanitary authorities. Copyright © 2012 Elsevier Masson SAS. All rights reserved.

  4. Species Diversity Effects on Productivity, Persistence and Quality of Multispecies Swards in a Four-Year Experiment

    DEFF Research Database (Denmark)

    Jing, Jingying; Søegaard, Karen; Cong, Wen-Feng

    2017-01-01

    ), neutral detergent fiber (NDF) and crude protein (CP), and a lower concentration of ash than the 10-mix and 12-mix. Slurry application increased annual yield production by 10% and changed the botanical composition, increasing the proportion of grass and decreasing the proportion of legumes. Compared......Plant species diversity may benefit natural grassland productivity, but its effect in managed grassland systems is not well understood. A four-year multispecies grassland experiment was conducted to investigate the effect of species diversity±legumes and non-leguminous forbs±on productivity...... increased sward production and yield persistence under cutting regime. The 12-mix had the highest yield from the second year onwards and no statistically significant yield reduction over four years, while annual yields in the 3-mix and 10-mix decreased significantly with increasing grassland age. The higher...

  5. Bacterial Pathogens and Antimicrobial Resistance Patterns in Pediatric Urinary Tract Infections: A Four-Year Surveillance Study (2009–2012)

    OpenAIRE

    Mirsoleymani, Seyed Reza; Salimi, Morteza; Shareghi Brojeni, Masoud; Ranjbar, Masoud; Mehtarpoor, Mojtaba

    2014-01-01

    The aims of this study were to assess the common bacterial microorganisms causing UTI and their antimicrobial resistance patterns in Bandar Abbas (Southern Iran) during a four-year period. In this retrospective study, samples with a colony count of ≥105 CFU/mL bacteria were considered positive; for these samples, the bacteria were identified, and the profile of antibiotic susceptibility was characterized. From the 19223 samples analyzed, 1513 (7.87%) were positive for bacterial infection. UTI...

  6. President Vladimir Putin’s Third Four-Year Term: Contradictory Outcomes – an Expec ted Result

    Directory of Open Access Journals (Sweden)

    Vladimir Aleksandrovich Ilyin

    2016-05-01

    Full Text Available In mid-March, Vladimir Putin’s third four-year presidency came to an end. If the term of office of the head of state were not prolonged in 2008, then the seventh presidential election would take place today, and therefore, a critical question arises: “What has been done and what needs to be done?... What does the President have in store for the people?”

  7. Modifiable risk factors predicting major depressive disorder at four year follow-up: a decision tree approach.

    Science.gov (United States)

    Batterham, Philip J; Christensen, Helen; Mackinnon, Andrew J

    2009-11-22

    Relative to physical health conditions such as cardiovascular disease, little is known about risk factors that predict the prevalence of depression. The present study investigates the expected effects of a reduction of these risks over time, using the decision tree method favoured in assessing cardiovascular disease risk. The PATH through Life cohort was used for the study, comprising 2,105 20-24 year olds, 2,323 40-44 year olds and 2,177 60-64 year olds sampled from the community in the Canberra region, Australia. A decision tree methodology was used to predict the presence of major depressive disorder after four years of follow-up. The decision tree was compared with a logistic regression analysis using ROC curves. The decision tree was found to distinguish and delineate a wide range of risk profiles. Previous depressive symptoms were most highly predictive of depression after four years, however, modifiable risk factors such as substance use and employment status played significant roles in assessing the risk of depression. The decision tree was found to have better sensitivity and specificity than a logistic regression using identical predictors. The decision tree method was useful in assessing the risk of major depressive disorder over four years. Application of the model to the development of a predictive tool for tailored interventions is discussed.

  8. Foresight beyond the very next event: Four-year-olds can link past and deferred future episodes

    Directory of Open Access Journals (Sweden)

    Jonathan eRedshaw

    2013-07-01

    Full Text Available Previous experiments have demonstrated that by four years of age children can use information from a past episode to solve a problem for the very next future episode. However, it remained unclear whether four-year-olds can similarly use such information to solve a problem for a more removed future episode that is not of immediate concern. In the current study we introduced four-year-olds to problems in one room before taking them to another room and distracting them for 15 minutes. The children were then offered a choice of items to place into a bucket that was to be taken back to the first room when a five-minute sand-timer had completed a cycle. Across two conceptually distinct domains, the children placed the item that could solve the deferred future problem above chance level. This result demonstrates that by 48 months many children can recall a problem from the past and act in the present to solve that problem for a deferred future episode. We discuss implications for theories about the nature of episodic foresight.

  9. Differentiating maternal fatigue and depressive symptoms at six months and four years post partum: Considerations for assessment, diagnosis and intervention.

    Science.gov (United States)

    Giallo, Rebecca; Gartland, Deirdre; Woolhouse, Hannah; Brown, Stephanie

    2015-02-01

    fatigue and depressive symptoms are common among women in the postpartum period, and it has been proposed that fatigue is a risk factor for later depression. To progress this research, there is a need to clarify the conceptual and measurement issue of whether these two sets of symptoms are distinct constructs. There is also a need to determine whether they are distinct constructs beyond the postnatal period. The aim of the study was to assess the construct and discriminant validity of fatigue and depressive symptoms as measured by the SF-36 Vitality subscale (SF-36) and the Edinburgh Postnatal Depression Scale (EPDS) at six months and at four years post partum. data from over 1000 women participating in the Maternal Health Study, a longitudinal study of women׳s physical and psychological health and recovery after childbirth were used. confirmatory factor analysis revealed a two-factor model of fatigue and depressive symptoms represented as distinct but related constructs was a better fit to the data than a one-factor model of fatigue and depression sharing the same underlying construct at both six months and four years post partum. this study provides empirical evidence that maternal fatigue and depression in the first year after having a baby and at four years post partum are best understood as separate psychological constructs or experiences. The findings have important implications for clinical practice, in particular underlining the importance of differentiating tiredness from depression. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Reduction in transmission of Schistosoma mansoni by a four-year focal mollusciciding programme against Biomphalaria glabrata in Saint Lucia.

    Science.gov (United States)

    Prentice, M A; Jordan, P; Bartholomew, R K; Grist, E

    1981-01-01

    The effect of transmission of Schistosoma mansoni of a focal snail control programme was investigated over four years amongst approximately 1250 people living in five communities in the steep-sided Soufriere river valley, St. Lucia, West Indies. Bayer 6076 was applied from constant flow drip cans to 12 stream sections at a target dose of 8 mg/litre clonitralide every four weeks. Only proven and potential transmission sites were treated; marsh habitats, where Biomphalaria glabrata were widespread, were ignored. In the stream snail numbers were reduced by 94% in the first year and by 100% thereafter. Incidence of new S. mansoni infections amongst children fell from 18% in the last year before control to 6% and 9% after three and four years respectively. Amongst children and adults in the four years of control the conversion/reversion ratio declined leading to a lowering of the over-all prevalence from 40% to 22%. Parasitologically the results were similar to those of a previously evaluated area-wide mollusciciding programme. The mean annual cost per person protected was US $2.60. This figure is atypically high because the topography of the area severely limited the population size.

  11. Modifiable risk factors predicting major depressive disorder at four year follow-up: a decision tree approach

    Directory of Open Access Journals (Sweden)

    Christensen Helen

    2009-11-01

    Full Text Available Abstract Background Relative to physical health conditions such as cardiovascular disease, little is known about risk factors that predict the prevalence of depression. The present study investigates the expected effects of a reduction of these risks over time, using the decision tree method favoured in assessing cardiovascular disease risk. Methods The PATH through Life cohort was used for the study, comprising 2,105 20-24 year olds, 2,323 40-44 year olds and 2,177 60-64 year olds sampled from the community in the Canberra region, Australia. A decision tree methodology was used to predict the presence of major depressive disorder after four years of follow-up. The decision tree was compared with a logistic regression analysis using ROC curves. Results The decision tree was found to distinguish and delineate a wide range of risk profiles. Previous depressive symptoms were most highly predictive of depression after four years, however, modifiable risk factors such as substance use and employment status played significant roles in assessing the risk of depression. The decision tree was found to have better sensitivity and specificity than a logistic regression using identical predictors. Conclusion The decision tree method was useful in assessing the risk of major depressive disorder over four years. Application of the model to the development of a predictive tool for tailored interventions is discussed.

  12. Chondrocyte activity is increased in psoriatic arthritis and axial spondyloarthritis

    DEFF Research Database (Denmark)

    Gudmann, Natasja Stæhr; Munk, Heidi Lausten; Christensen, Anne Friesgaard

    2016-01-01

    . There is a need for biomarkers reflecting core disease pathways for diagnosis and disease mapping. Pro-C2 reflects mature cartilage collagen type IIB formation, while C-Col10 represents turnover of type X collagen, which is exclusively expressed by hypertrophic chondrocytes. The objectives of this study were......SpA undergoing TNFi treatment may reflect that hypertrophic chondrocytes in axSpA are targeted by TNFi. ROC curve analysis showed a diagnostic potential for Pro-C2 in axSpA and PsA....

  13. Four years of crusade!

    Directory of Open Access Journals (Sweden)

    Adilson Beatriz

    2012-04-01

    Full Text Available Orbital - The Electronic Journal of Chemistry is entering its 4th year of existence which is a reason to celebrate. A great work has been devoted to preserve a scientific journal of international reputation. This work was not always without mistakes or any trouble. However, these very adversities stimulate additional efforts in order to overcome our errors and, continuing with our serious proposal of editing a journal which scientists from all over the world can submit online their original works manuscripts to be published in peer review and open-access style. Beginning this year, Orbital was classified by the Chemistry Coordination Area on the B5 rank of CAPES Qualis (Brazil and, leveled as B4 by Coordination Areas of Materials and Engineering II. These important details demonstrate that Orbital is progressively attaining the acknowledgment of the Brazilian scientific community. The recognition by CAPES Qualis is crucial as the system evaluates the generation of knowledge in the Brazilian universities and the method by which this same knowledge disseminated to society. Not all these were possible without endeavoring of editors and volunteer assistance of advisors from several national and international institutions. Recently, the young Professors Kleber Thiago de Oliveira (UFSCAR, Grégoire Demetz (USP-RP and Amilcar Machulek Júnior (UFMS have joined us to share the idea and philosophy of Orbital by accepting our invitation to be associated editors. The tasks and challenges are enormous and, we are sure that new participants will support us on the way to achieve indexation in new Databases systems as SCOPUS, ISI and Scielo. Consequently, the international success of this electronic journal will be assured. We sincerely hope that in the academic Brazilian scenario, Orbital can figure as a solid bridge to interchange real creative science that is happening in various locations around the world.

  14. Four years of HELAS

    Science.gov (United States)

    Roth, M.; Lühe, O. v. d.; Aerts, C.; Christensen-Dalsgaard, J.; Corbard, T.; Daszyńska-Daszkiewicz, J.; Di Mauro, M. P.; Gizon, L.; Jiménez-Reyes, S.; Monteiro, M. J. P. F. G.; Pallé, P. L.; Thompson, M. J.

    2010-12-01

    The European Coordination Action on HELio- and ASteroseismology (HELAS) has completed its fourth and final year of initial funding by the European Commission. Set up as a network which combines solar and stellar physics communities in the important and vigorously evolving field of seismology, HELAS has been able to coordinate the efforts of European astronomers with remarkable success. Four large international conferences including the HELAS-IV conference on Lanzarote as well as many workshops were organized with a substantial contribution from HELAS. About a dozen workshops, addressing specialized questions in global and local helioseismology and asteroseismology were entirely organized by HELAS. Data analysis tools to prepare the European communities for the upcoming influx of data from new missions have been prepared, tested and demonstrated. Lecture notes and outreach material have been assembled and prepared for general access. As a result, HELAS has an important impact on the scientific output of the astrophysics seismology communities and significantly increased the visibility of European research in this field. This paper summarizes the activities and accomplishments of HELAS.

  15. [Visual outcomes four years after small incision lenticule extraction (SMILE) surgery on highly myopic eyes [French version

    Science.gov (United States)

    Burazovitch, J; Naguzeswski, D; Beuste, T; Guillard, M

    2017-09-01

    To determine whether the visual outcomes of the refractive surgery technique, small incision lenticule extraction (SMILE), are stable, effective, and predictable for high myopia over a four-year period. This is a retrospective study. The data were collected between March 2012 and July 2016. Two hundred and forty-eight patients participated in the study; that is, 496 eyes: 140 eyes of 70 patients (52 women/18 men) were classified in the highly myopic group (refraction measured in spherical equivalent [RMSE]>-6D), and 356 eyes of 178 patients (98 women/80 men) in the control group (RMSE≤-6D). Follow-up tests were conducted immediately post-operatively (D+1), after three months, after one year, and after four years. Refraction, uncorrected visual acuity (UCVA), and best visual corrected acuity (BCVA) were measured. The highly myopic group (HMG) contained more women, and astigmatism was higher for this group than for the control group (CG). These are BCVA, refractive stability, the index of safety (SI: BCVA preoperatively D+1/BCVA postoperatively), and predictability (the percentage of eyes within±0.5 D of the target). In both groups, UCVA was better after the fourth year than it was immediately after the procedure (HMG: P=0.001; CG: P=0.001). Although it differed at one year (P=0.01), the groups' refractive stability tended to converge over four years (P=0.138). Both groups' SI was identical in the four follow-up tests (P=0.734 at D+1; P=0.07 at M+1; P=0.160 at M3 and Y1; and P=0.274 at Y4). For the HMG, SI stability was attained after three months (1.00±0.1); whereas it was attained after one month (0.91±0.11) for the CG. Four years after the surgery, we observed that 87 % of the operated eyes in the HMG were within 0.5 D of the target. SMILE is a good refractive surgery technique for treating high myopia. It yields stable, safe, effective, and predictable results over four years. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  16. Comparative study of the chondrogenic potential of human bone marrow stromal cells, neonatal chondrocytes and adult chondrocytes

    International Nuclear Information System (INIS)

    Saha, Sushmita; Kirkham, Jennifer; Wood, David; Curran, Stephen; Yang, Xuebin

    2010-01-01

    Research highlights: → This study has characterised three different cell types under conditions similar to those used for autologous chondrocyte implantation (ACI) for applications in cartilage repair/regeneration. → Compared for the first time the chondrogenic potential of neonatal chondrocytes with human bone marrow stromal cells (HBMSCs) and adult chondrocytes. → Demonstrated that adult chondrocytes hold greatest potential for use in ACI based on their higher proliferation rates, lower alkaline phosphatise activity and enhanced expression of chondrogenic genes. → Demonstrated the need for chondroinduction as a necessary pre-requisite to efficient chondrogenesis in vitro and, by extrapolation, for cell based therapy (e.g. ACI or cartilage tissue engineering). -- Abstract: Cartilage tissue engineering is still a major clinical challenge with optimisation of a suitable source of cells for cartilage repair/regeneration not yet fully addressed. The aims of this study were to compare and contrast the differences in chondrogenic behaviour between human bone marrow stromal cells (HBMSCs), human neonatal and adult chondrocytes to further our understanding of chondroinduction relative to cell maturity and to identify factors that promote chondrogenesis and maintain functional homoeostasis. Cells were cultured in monolayer in either chondrogenic or basal medium, recapitulating procedures used in existing clinical procedures for cell-based therapies. Cell doubling time, morphology and alkaline phosphatase specific activity (ALPSA) were determined at different time points. Expression of chondrogenic markers (SOX9, ACAN and COL2A1) was compared via real time polymerase chain reaction. Amongst the three cell types studied, HBMSCs had the highest ALPSA in basal culture and lowest ALPSA in chondrogenic media. Neonatal chondrocytes were the most proliferative and adult chondrocytes had the lowest ALPSA in basal media. Gene expression analysis revealed a difference in the

  17. Comparative study of the chondrogenic potential of human bone marrow stromal cells, neonatal chondrocytes and adult chondrocytes

    Energy Technology Data Exchange (ETDEWEB)

    Saha, Sushmita [Biomaterials and Tissue Engineering Group, Leeds Dental Institute, University of Leeds, LS29LU (United Kingdom); Kirkham, Jennifer [Biomineralisation Group, Leeds Dental Institute, University of Leeds, LS29LU (United Kingdom); NIHR Leeds Musculoskeletal Biomedical Research Unit, University of Leeds, Chapel Allerton Hospital, Leeds LS74SA (United Kingdom); Wood, David [Biomaterials and Tissue Engineering Group, Leeds Dental Institute, University of Leeds, LS29LU (United Kingdom); Curran, Stephen [Smith and Nephew Research Centre, YO105DF (United Kingdom); Yang, Xuebin, E-mail: X.B.Yang@leeds.ac.uk [Biomaterials and Tissue Engineering Group, Leeds Dental Institute, University of Leeds, LS29LU (United Kingdom); NIHR Leeds Musculoskeletal Biomedical Research Unit, University of Leeds, Chapel Allerton Hospital, Leeds LS74SA (United Kingdom)

    2010-10-22

    Research highlights: {yields} This study has characterised three different cell types under conditions similar to those used for autologous chondrocyte implantation (ACI) for applications in cartilage repair/regeneration. {yields} Compared for the first time the chondrogenic potential of neonatal chondrocytes with human bone marrow stromal cells (HBMSCs) and adult chondrocytes. {yields} Demonstrated that adult chondrocytes hold greatest potential for use in ACI based on their higher proliferation rates, lower alkaline phosphatise activity and enhanced expression of chondrogenic genes. {yields} Demonstrated the need for chondroinduction as a necessary pre-requisite to efficient chondrogenesis in vitro and, by extrapolation, for cell based therapy (e.g. ACI or cartilage tissue engineering). -- Abstract: Cartilage tissue engineering is still a major clinical challenge with optimisation of a suitable source of cells for cartilage repair/regeneration not yet fully addressed. The aims of this study were to compare and contrast the differences in chondrogenic behaviour between human bone marrow stromal cells (HBMSCs), human neonatal and adult chondrocytes to further our understanding of chondroinduction relative to cell maturity and to identify factors that promote chondrogenesis and maintain functional homoeostasis. Cells were cultured in monolayer in either chondrogenic or basal medium, recapitulating procedures used in existing clinical procedures for cell-based therapies. Cell doubling time, morphology and alkaline phosphatase specific activity (ALPSA) were determined at different time points. Expression of chondrogenic markers (SOX9, ACAN and COL2A1) was compared via real time polymerase chain reaction. Amongst the three cell types studied, HBMSCs had the highest ALPSA in basal culture and lowest ALPSA in chondrogenic media. Neonatal chondrocytes were the most proliferative and adult chondrocytes had the lowest ALPSA in basal media. Gene expression analysis revealed

  18. Clinical Outcomes of Characterized Chondrocyte Implantation

    Science.gov (United States)

    Huylebroek, José; Van Der Bauwhede, Jan; Saris, Daniël; Veeckman, Geert; Bobic, Vladimir; Victor, Jan; Almqvist, Karl Fredrik; Verdonk, Peter; Fortems, Yves; Van Lommel, Nel; Haazen, Ludo

    2012-01-01

    Objective: To assess the clinical outcome of patients treated with autologous chondrocyte implantation using ChondroCelect in daily practice. Methods: The study is a cross-sectional analysis of an open-label, noninterventional cohort. The setting was a compassionate use program, involving 43 orthopaedic centers in 7 European countries. The participants were patients treated with ChondroCelect between October 13, 2004 and July 2, 2008. The measurements used were Clinical Global Impression–Improvement and –Efficacy and solicited adverse event reports. Results: Safety data were collected from 334 patients (90.3%), and effectiveness data were from 282 (76.2%) of the 370 patients treated. Mean age at baseline was 33.6 years (range, 12-57 years), 57% were male, and mean body mass index was 25 kg/m2. Mean follow-up was 2.2 years (range, 0.4-4.1 years). A femoral condyle lesion was reported in 66% (288/379) and a patellar lesion in 19% (84/379). Mean lesion size was 3.5 cm2; a collagen membrane was used in 92.4% (328/355). A therapeutic effect was reported in 89% (234/264) of patients overall and in 87% (40/46) of patellar lesion patients. Rates of much or very much improved patients were similar in patients with short- (18 months: 68% [70/103]) (P = 0.68) and were independent of lesion size (>4 cm2: 75.5% [37/49]; ≤4 cm2: 67.7% [111/164]) (P = 0.38). Adverse events were similar to those reported in the randomized trial with the same product, with more arthrofibrosis, more reduced joint mobility, and more crepitations reported in patellar lesions. Overall, less cartilage hypertrophy was noted, probably due to the use of a biological membrane cover. Conclusions: Implantation of ChondroCelect appeared to result in a positive benefit/risk ratio when used in an unselected heterogenous population, irrespective of the follow-up period, lesion size, and type of lesion treated. PMID:26069630

  19. Murine pluripotent stem cells derived scaffold-free cartilage grafts from a micro-cavitary hydrogel platform.

    Science.gov (United States)

    He, Pengfei; Fu, Jiayin; Wang, Dong-An

    2016-04-15

    By means of appropriate cell type and scaffold, tissue-engineering approaches aim to construct grafts for cartilage repair. Pluripotent stem cells especially induced pluripotent stem cells (iPSCs) are of promising cell candidates due to the pluripotent plasticity and abundant cell source. We explored three dimensional (3D) culture and chondrogenesis of murine iPSCs (miPSCs) on an alginate-based micro-cavity hydrogel (MCG) platform in pursuit of fabricating synthetic-scaffold-free cartilage grafts. Murine embryonic stem cells (mESCs) were employed in parallel as the control. Chondrogenesis was fulfilled using a consecutive protocol via mesoderm differentiation followed by chondrogenic differentiation; subsequently, miPSC and mESC-seeded constructs were further respectively cultured in chondrocyte culture (CC) medium. Alginate phase in the constructs was then removed to generate a graft only comprised of induced chondrocytic cells and cartilaginous extracellular matrix (ECMs). We found that from the mESC-seeded constructs, formation of intact grafts could be achieved in greater sizes with relatively fewer chondrocytic cells and abundant ECMs; from miPSC-seeded constructs, relatively smaller sized cartilaginous grafts could be formed by cells with chondrocytic phenotype wrapped by abundant and better assembled collagen type II. This study demonstrated successful creation of pluripotent stem cells-derived cartilage/chondroid graft from a 3D MCG interim platform. By the support of materials and methodologies established from this study, particularly given the autologous availability of iPSCs, engineered autologous cartilage engraftment may be potentially fulfilled without relying on the limited and invasive autologous chondrocytes acquisition. In this study, we explored chondrogenic differentiation of pluripotent stem cells on a 3D micro-cavitary hydrogel interim platform and creation of pluripotent stem cells-derived cartilage/chondroid graft via a consecutive

  20. Widespread epigenomic, transcriptomic and proteomic differences between hip osteophytic and articular chondrocytes in osteoarthritis.

    Science.gov (United States)

    Steinberg, Julia; Brooks, Roger A; Southam, Lorraine; Bhatnagar, Sahir; Roumeliotis, Theodoros I; Hatzikotoulas, Konstantinos; Zengini, Eleni; Wilkinson, J Mark; Choudhary, Jyoti S; McCaskie, Andrew W; Zeggini, Eleftheria

    2018-05-08

    To identify molecular differences between chondrocytes from osteophytic and articular cartilage tissue from OA patients. We investigated genes and pathways by combining genome-wide DNA methylation, RNA sequencing and quantitative proteomics in isolated primary chondrocytes from the cartilaginous layer of osteophytes and matched areas of low- and high-grade articular cartilage across nine patients with OA undergoing hip replacement surgery. Chondrocytes from osteophytic cartilage showed widespread differences to low-grade articular cartilage chondrocytes. These differences were similar to, but more pronounced than, differences between chondrocytes from osteophytic and high-grade articular cartilage, and more pronounced than differences between high- and low-grade articular cartilage. We identified 56 genes with significant differences between osteophytic chondrocytes and low-grade articular cartilage chondrocytes on all three omics levels. Several of these genes have known roles in OA, including ALDH1A2 and cartilage oligomeric matrix protein, which have functional genetic variants associated with OA from genome-wide association studies. An integrative gene ontology enrichment analysis showed that differences between osteophytic and low-grade articular cartilage chondrocytes are associated with extracellular matrix organization, skeletal system development, platelet aggregation and regulation of ERK1 and ERK2 cascade. We present a first comprehensive view of the molecular landscape of chondrocytes from osteophytic cartilage as compared with articular cartilage chondrocytes from the same joints in OA. We found robust changes at genes relevant to chondrocyte function, providing insight into biological processes involved in osteophyte development and thus OA progression.

  1. Low‑dose halofuginone inhibits the synthesis of type I collagen without influencing type II collagen in the extracellular matrix of chondrocytes.

    Science.gov (United States)

    Li, Zeng; Fei, Hao; Wang, Zhen; Zhu, Tianyi

    2017-09-01

    Full‑thickness and large area defects of articular cartilage are unable to completely repair themselves and require surgical intervention, including microfracture, autologous or allogeneic osteochondral grafts, and autologous chondrocyte implantation. A large proportion of regenerative cartilage exists as fibrocartilage, which is unable to withstand impacts in the same way as native hyaline cartilage, owing to excess synthesis of type I collagen in the matrix. The present study demonstrated that low‑dose halofuginone (HF), a plant alkaloid isolated from Dichroa febrifuga, may inhibit the synthesis of type I collagen without influencing type II collagen in the extracellular matrix of chondrocytes. In addition, HF was revealed to inhibit the phosphorylation of mothers against decapentaplegic homolog (Smad)2/3 and promoted Smad7 expression, as well as decrease the synthesis of type I collagen synthesis. Results from the present study indicated that HF treatment suppressed the synthesis of type I collagen by inhibiting the transforming growth factor‑β signaling pathway in chondrocytes. These results may provide an alternative solution to the problems associated with fibrocartilage, and convert fibrocartilage into hyaline cartilage at the mid‑early stages of cartilage regeneration. HF may additionally be used to improve monolayer expansion or 3D cultures of seed cells for the tissue engineering of cartilage.

  2. Grafting and curing

    International Nuclear Information System (INIS)

    Garnett, J.L.; Loo-Teck Ng; Visay Viengkhou

    1998-01-01

    Progress in radiation grafting and curing is briefly reviewed. The two processes are shown to be mechanistically related. The parameters influencing yields are examined particularly for grafting. For ionising radiation grafting systems (EB and gamma ray) these include solvents, substrate and monomer structure, dose and dose-rate, temperature and more recently role of additives. In addition, for UV grafting, the significance of photoinitiators is discussed. Current applications of radiation grafting and curing are outlined. The recent development of photoinitiator free grafting and curing is examined as well as the potential for the new excimer laser sources. The future application of both grafting and curing is considered, especially the significance of the occurrence of concurrent grafting during cure and its relevance in environmental considerations

  3. Skin graft - slideshow

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/presentations/100100.htm Skin graft - series—Normal anatomy To use the sharing features ... entire body, and acts as a protective barrier. Skin grafts may be recommended for: Extensive wounds Burns Specific ...

  4. Association of Circulating 25(OHD and Lower Urinary Tract Symptoms: A Four-Year Prospective Study among Elderly Chinese Men

    Directory of Open Access Journals (Sweden)

    Zhao-Min Liu

    2016-05-01

    Full Text Available The role of vitamin D in relation to lower urinary tract symptoms (LUTS remains inconclusive. This four-year longitudinal study aims to explore the association of circulating 25(OHD and LUTS in elderly Chinese men. Two thousand Chinese men aged 65 and older were recruited from a local community, of which 1998 (99.9% at baseline and 1564 (78.2% at four-year follow-up reported data on LUTS, and 988 of the randomly chosen subpopulation were assayed for serum 25(OHD by radioimmunoassay at baseline. LUTS were evaluated by a validated International Prostate Symptoms Scale (IPSS. Data on demographic characteristics, lifestyle factors, health, and medications were collected. Serum parathyroid and sex steroid hormones and genotypes of vitamin D receptors were assayed. The association of serum 25(OHD and LUTS was examined by using multivariable regression models. Serum 25(OHD was not significantly associated with the changes of IPSS or the risk of LUTS in overall participants. However, among men with 25(OHD ≤ 60 nmol/L, each 10 nmol/L increase of 25(OHD over 0 nmol/L was significantly associated with 1.3 lower points of IPSS or a 51.6% decreased risk for moderate/severe LUTS four years later. Adjustment for serum androstenedione (p = 0.019 and dehydropiandrosterone (p = 0.037 attenuated the associations. Our study suggested that among individuals with low vitamin D status, the increase of the 25(OHD level may be associated with a lowered risk of LUTS.

  5. The role of PTHrP in chondrocyte differentiation

    NARCIS (Netherlands)

    Hoogendam, Jakomijn

    2006-01-01

    Longitudinal growth is the key characteristic that distinguishes children from adults. Growth is regulated in the growth plates, which are layers of cartilage located at the ends of the long bones. The cartilage cells are called chondrocytes and go through a coordinated program of proliferation,

  6. Effect of donor age on DNA repair by articular chondrocytes

    International Nuclear Information System (INIS)

    Lipman, J.M.

    1986-01-01

    The hypothesis that aging of articular chondrocytes at a cellular level results from loss of DNA repair capability was studied by two different measures: unscheduled DNA synthesis (UDS) and O 6 -methylguanine acceptor protein (MGAP) activity. UDS following damage by 254 nm ultraviolet irradiation (20J/m 2 ) was examined in intact articular cartilage from rabbits of different ages. Semiconservative DNA synthesis was suppressed with hydroxurea and repair followed by the incorporation of [ 3 H]-thymidine ([ 3 H]-dThd). After repair the cartilage was digested in proteinase K (0.5mg/ml) with dodecyl sodium sulfate (0.2%) and DNA determined with Hoechst 33258 dye. UDS (dpm [ 3 H]-dThd/μg DNA) was greater in articular cartilage from 3- than 39-month-old rabbits. MGAP was studied in cell extracts of cultured human and rabbit chondrocytes by transfer of [ 3 H] O 6 -methyl groups from exogenous DNA to protein. It was significantly less in rabbit than in human cells on a per protein or DNA basis. There was no decline in this activity in human chondrocytes from newborn to 60 years of age; and rabbits from 3- to 36-months-old. The data indicate that in the two different repair mechanisms, age differences are found with resting but not dividing chondrocytes

  7. Mechanical confinement regulates cartilage matrix formation by chondrocytes

    Science.gov (United States)

    Lee, Hong-Pyo; Gu, Luo; Mooney, David J.; Levenston, Marc E.; Chaudhuri, Ovijit

    2017-12-01

    Cartilage tissue equivalents formed from hydrogels containing chondrocytes could provide a solution for replacing damaged cartilage. Previous approaches have often utilized elastic hydrogels. However, elastic stresses may restrict cartilage matrix formation and alter the chondrocyte phenotype. Here we investigated the use of viscoelastic hydrogels, in which stresses are relaxed over time and which exhibit creep, for three-dimensional (3D) culture of chondrocytes. We found that faster relaxation promoted a striking increase in the volume of interconnected cartilage matrix formed by chondrocytes. In slower relaxing gels, restriction of cell volume expansion by elastic stresses led to increased secretion of IL-1β, which in turn drove strong up-regulation of genes associated with cartilage degradation and cell death. As no cell-adhesion ligands are presented by the hydrogels, these results reveal cell sensing of cell volume confinement as an adhesion-independent mechanism of mechanotransduction in 3D culture, and highlight stress relaxation as a key design parameter for cartilage tissue engineering.

  8. Young woman with a four-year history of epilepsy and progressive focal cortical atrophy — What is the diagnosis?

    Directory of Open Access Journals (Sweden)

    S. Pati

    2014-01-01

    Full Text Available The pathogenesis of disease progression in drug-refractory epilepsy is poorly understood. We report the case of a young woman with a four-year history of epilepsy that progressed rapidly as evidenced by the development of progressive focal cortical atrophy. She underwent biopsy that showed perinatal ischemia and a prominent inflammatory response, including T-cell infiltration and microglial activation. There was no consensus reached on the final diagnosis although the hypothesis of dual pathology (adult variant of Rasmussen's encephalitis and perinatal stroke was considered. The possible role of inflammation in the progression of epilepsy caused by a “static” lesion (perinatal stroke is discussed.

  9. Cartilage Morphological and Histological Findings After Reconstruction of the Glenoid With an Iliac Crest Bone Graft.

    Science.gov (United States)

    Auffarth, Alexander; Resch, Herbert; Matis, Nicholas; Hudelmaier, Martin; Wirth, Wolfgang; Forstner, Rosemarie; Neureiter, Daniel; Traweger, Andreas; Moroder, Philipp

    2018-04-01

    The J-bone graft is presumably representative of iliac crest bone grafts in general and allows anatomic glenoid reconstruction in cases of bone defects due to recurrent traumatic anterior shoulder dislocations. As a side effect, these grafts have been observed to be covered by some soft, cartilage-like tissue when arthroscopy has been indicated after such procedures. To evaluate the soft tissue covering of J-bone grafts by use of magnetic resonance imaging (MRI) and histological analysis. Case series; Level of evidence, 4. Patients underwent MRI at 1 year after the J-bone graft procedures. Radiological data were digitally processed and evaluated by segmentation of axial images. Independent from the MRI analysis, 2 biopsy specimens of J-bone grafts were harvested for descriptive histological analysis. Segmentation of the images revealed that all grafts were covered by soft tissue. This layer had an average thickness of 0.87 mm compared with 1.96 mm at the adjacent native glenoid. Of the 2 biopsy specimens, one exhibited evident hyaline-like cartilage and the other presented patches of chondrocytes embedded in a glycosaminoglycan-rich extracellular matrix. J-bone grafts are covered by soft tissue that can differentiate into fibrous and potentially hyaline cartilage. This feature may prove beneficial for delaying the onset of dislocation arthropathy of the shoulder.

  10. Influence of cell printing on biological characters of chondrocytes.

    Science.gov (United States)

    Qu, Miao; Gao, Xiaoyan; Hou, Yikang; Shen, Congcong; Xu, Yourong; Zhu, Ming; Wang, Hengjian; Xu, Haisong; Chai, Gang; Zhang, Yan

    2015-01-01

    To establish a two-dimensional biological printing technique of chondrocytes and compare the difference of related biological characters between printed chondrocytes and unprinted cells so as to control the cell transfer process and keep cell viability after printing. Primary chondrocytes were obtained from human mature and fetal cartilage tissues and then were regularly sub-cultured to harvest cells at passage 2 (P2), which were adjusted to the single cell suspension at a density of 1×10(6)/mL. The experiment was divided into 2 groups: experimental group P2 chondrocytes were transferred by rapid prototype biological printer (driving voltage value 50 V, interval in x-axis 300 μm, interval in y-axis 1500 μm). Afterwards Live/Dead viability Kit and flow cytometry were respectively adopted to detect cell viability; CCK-8 Kit was adopted to detect cell proliferation viability; immunocytochemistry, immunofluorescence and RT-PCR was employed to identify related markers of chondrocytes; control group steps were the same as the printing group except that cell suspension received no printing. Fluorescence microscopy and flow cytometry analyses showed that there was no significant difference between experimental group and control group in terms of cell viability. After 7-day in vitro culture, control group exhibited higher O.D values than experimental group from 2nd day to 7th day but there was no distinct difference between these two groups (P>0.05). Inverted microscope observation demonstrated that the morphology of these two groups had no significant difference either. Similarly, Immunocytochemistry, immunofluorescence and RT-PCR assays also showed that there was no significant difference in the protein and gene expression of type II collagen and aggrecan between these two groups (P>0.05). Conclusion Cell printing has no distinctly negative effect on cell vitality, proliferation and phenotype of chondrocytes. Biological printing technique may provide a novel approach

  11. Bone grafts in dentistry

    Directory of Open Access Journals (Sweden)

    Prasanna Kumar

    2013-01-01

    Full Text Available Bone grafts are used as a filler and scaffold to facilitate bone formation and promote wound healing. These grafts are bioresorbable and have no antigen-antibody reaction. These bone grafts act as a mineral reservoir which induces new bone formation.

  12. RAGE, receptor of advanced glycation endoproducts, negatively regulates chondrocytes differentiation.

    Directory of Open Access Journals (Sweden)

    Tatsuya Kosaka

    Full Text Available RAGE, receptor for advanced glycation endoproducts (AGE, has been characterized as an activator of osteoclastgenesis. However, whether RAGE directly regulates chondrocyte proliferation and differentiation is unclear. Here, we show that RAGE has an inhibitory role in chondrocyte differentiation. RAGE expression was observed in chondrocytes from the prehypertrophic to hypertrophic regions. In cultured cells, overexpression of RAGE or dominant-negative-RAGE (DN-RAGE demonstrated that RAGE inhibited cartilaginous matrix production, while DN-RAGE promoted production. Additionally, RAGE regulated Ihh and Col10a1 negatively but upregulated PTHrP receptor. Ihh promoter analysis and real-time PCR analysis suggested that downregulation of Cdxs was the key for RAGE-induced inhibition of chondrocyte differentiation. Overexpression of the NF-κB inhibitor I-κB-SR inhibited RAGE-induced NF-κB activation, but did not influence inhibition of cartilaginous matrix production by RAGE. The inhibitory action of RAGE was restored by the Rho family GTPases inhibitor Toxin B. Furthermore, inhibitory action on Ihh, Col10a1 and Cdxs was reproduced by constitutively active forms, L63RhoA, L61Rac, and L61Cdc42, but not by I-κB-SR. Cdx1 induced Ihh and Col10a1 expressions and directly interacted with Ihh promoter. Retinoic acid (RA partially rescued the inhibitory action of RAGE. These data combined suggests that RAGE negatively regulates chondrocyte differentiation at the prehypertrophic stage by modulating NF-κB-independent and Rho family GTPases-dependent mechanisms.

  13. Nanosized fibers' effect on adult human articular chondrocytes behavior

    International Nuclear Information System (INIS)

    Stenhamre, Hanna; Thorvaldsson, Anna; Enochson, Lars; Walkenström, Pernilla; Lindahl, Anders; Brittberg, Mats; Gatenholm, Paul

    2013-01-01

    Tissue engineering with chondrogenic cell based therapies is an expanding field with the intention of treating cartilage defects. It has been suggested that scaffolds used in cartilage tissue engineering influence cellular behavior and thus the long-term clinical outcome. The objective of this study was to assess whether chondrocyte attachment, proliferation and post-expansion re-differentiation could be influenced by the size of the fibers presented to the cells in a scaffold. Polylactic acid (PLA) scaffolds with different fiber morphologies were produced, i.e. microfiber (MS) scaffolds as well as nanofiber-coated microfiber scaffold (NMS). Adult human articular chondrocytes were cultured in the scaffolds in vitro up to 28 days, and the resulting constructs were assessed histologically, immunohistochemically, and biochemically. Attachment of cells and serum proteins to the scaffolds was affected by the architecture. The results point toward nano-patterning onto the microfibers influencing proliferation of the chondrocytes, and the overall 3D environment having a greater influence on the re-differentiation. In the efforts of finding the optimal scaffold for cartilage tissue engineering, studies as the current contribute to the knowledge of how to affect and control chondrocytes behavior. - Highlights: ► Chondrocyte behavior in nanofiber-coated microfiber versus microfiber scaffolds ► High porosity (> 90%) and large pore sizes (a few hundred μm) of nanofibrous scaffolds ► Proliferation enhanced by presence of nanofibers ► Differentiation not significantly affected ► Cell attachment improved in presence of both nanofibers and serum

  14. Population profiles associated with severe functioning difficulties and disability among two to four years old children in Mexico

    Directory of Open Access Journals (Sweden)

    Rosalba Rojas-Martínez

    2017-07-01

    Full Text Available Objective. To describe the prevalence and factors associated with severe child functioning difficulties and disability (CFD among two to four year old children in Mexico, and estimates the probability of presenting CFD based on specific population profiles. Materials and methods. The sample consists of 5 104 children who participated in the National Survey of Children and Women 2015 (ENIM. We used postestimation exploration by computing predicted values of CFD to interpret the logistic models for discrete combinations of the independent variables. Results. CFD prevalence is 2%,which means at least 130 000 two to four year-old children are at risk of experiencing severely limited participation in an unaccommodating environment. The probability of presenting CFD is dramatically higher in specific sub-groups of the population, in particular, male children of women with low education, who live in the poorest households. Conclusions. A significant proportion of Mexican children face important challenges due to functioning difficulties and disability. Public policies must be developed to accommodate the needs of these children and provide a proper environment for their development.

  15. Extension of general practice training from three to four years: experiences of a vocational training programme in Southern Ireland.

    LENUS (Irish Health Repository)

    Dowling, Stephanie

    2009-05-01

    The aim of this study was to evaluate the experiences of trainees taking part in an extended (four-year) general practice training programme introduced in the South Eastern region of the Republic of Ireland to replace the previous traditional (three-year) programme. In a qualitative design, eight homogeneous focus groups were held to determine the value of the additional year of training. The first cohort of trainees was interviewed towards the start and at the end of their fourth year. Trainees finishing the following year were also interviewed, as were graduates from the final three-year programme. GP trainers and the four members of the programme directing team comprised two further independent focus groups. Trainees reported that the integration of hospital posts and general practice attachments over the four years was particularly beneficial. The exposure to a variety of different general practices and the opportunity to take part in specialty clinics were considered extremely useful. The fourth year of training was felt to be less pressurised than previous years. Professional and personal development was enhanced; improved readiness to practise and confidence were noted. Perceived disadvantages of extended training included a lack of acknowledgment for doctors in their fourth year and excessive emphasis placed on research during the final year of training. The addition of an extra year of vocational training improves professional and personal development and changes the learning experience for doctors. Doctors felt more confident and ready to enter independent practice at the end of the fourth year of training.

  16. Four-year-olds' strategic allocation of resources: attempts to elicit reciprocation correlate negatively with spontaneous helping.

    Science.gov (United States)

    Kenward, Ben; Hellmer, Kahl; Winter, Lina Söderström; Eriksson, Malin

    2015-03-01

    Behaviour benefitting others (prosocial behaviour) can be motivated by self-interested strategic concerns as well as by genuine concern for others. Even in very young children such behaviour can be motivated by concern for others, but whether it can be strategically motivated by self-interest is currently less clear. Here, children had to distribute resources in a game in which a rich but not a poor recipient could reciprocate. From four years of age participants strategically favoured the rich recipient, but only when recipients had stated an intention to reciprocate. Six- and eight-year-olds distributed more equally. Children allocating strategically to the rich recipient were less likely to help when an adult needed assistance but was not in a position to immediately reciprocate, demonstrating consistent cross-task individual differences in the extent to which social behaviour is self- versus other-oriented even in early childhood. By four years of age children are capable of strategically allocating resources to others as a tool to advance their own self-interest. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Where to find a trustworthy source of information? four years of risk communication about a nuclear waste disposal

    International Nuclear Information System (INIS)

    Findahl, O.

    1998-01-01

    Full text of publication follows: in September 1997 there was a referendum, in a small local district in the north of Sweden, on the disposal of the swedish high intensive nuclear waste. A small majority of the 2500 voters stopped the examination of the conditions for a storage. Four years earlier the opinion against a local storage of nuclear waste had been much stronger but during the years that followed more people came to think that an examination of the conditions for a storage was OK; that will create new jobs, and what will come later we will see. The closer to the referendum the stronger this positive opinion became. They did not have any information problems. There were experts and information sources they could believe in. But the majority was still negative and they were dissatisfied. They could not find any trustworthy information. And though they in the end had prevented the localization of the nuclear waste storage to their commune they still felt that the decision had been taken high above their heads. Why is it like that? And what happened during these four years of experimentation in democratic risk communication? (author)

  18. Use of Preputial Skin as Cutaneous Graft after Nevus Excision

    Directory of Open Access Journals (Sweden)

    A. D'Alessio

    2010-01-01

    Full Text Available We report a four-year-old boy with a nevus covering all the plantar side of his second finger on the left foot. He was also affected by congenital phimosis. Surgical excision of the nevus was indicated, but the skin defect would have been too large to be directly closed. The foreskin was taken as a full-thickness skin graft to cover the cutaneous defect of the finger. The graft intake was favourable and provided a functional repair with good aesthetic characteristic.

  19. Viability of chondrocytes seeded onto a collagen I/III membrane for matrix-induced autologous chondrocyte implantation.

    Science.gov (United States)

    Hindle, Paul; Hall, Andrew C; Biant, Leela C

    2014-11-01

    Cell viability is crucial for effective cell-based cartilage repair. The aim of this study was to determine the effect of handling the membrane during matrix-induced autologous chondrocyte implantation surgery on the viability of implanted chondrocytes. Images were acquired under five conditions: (i) Pre-operative; (ii) Handled during surgery; (iii) Cut edge; (iv) Thumb pressure applied; (v) Heavily grasped with forceps. Live and dead cell stains were used. Images were obtained for cell counting and morphology. Mean cell density was 6.60 × 10(5) cells/cm(2) (5.74-7.11 × 10(5) ) in specimens that did not have significant trauma decreasing significantly in specimens that had been grasped with forceps (p < 0.001) or cut (p = 0.004). Cell viability on delivery grade membrane was 75.1%(72.4-77.8%). This dropped to 67.4%(64.1-69.7%) after handling (p = 0.002), 56.3%(51.5-61.6%) after being thumbed (p < 0.001) and 28.8%(24.7-31.2%) after crushing with forceps (p < 0.001). When cut with scissors there was a band of cell death approximately 275 µm in width where cell viability decreased to 13.7%(10.2-18.2%, p < 0.001). Higher magnification revealed cells without the typical rounded appearance of chondrocytes. We found that confocal laser-scanning microscope (CLSM) can be used to quantify and image the fine morphology of cells on a matrix-induced autologous chondrocyte implantation (MACI) membrane. Careful handling of the membrane is essential to minimise chondrocyte death during surgery. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  20. The inhibitory roles of Ihh downregulation on chondrocyte growth and differentiation.

    Science.gov (United States)

    Deng, Ang; Zhang, Hongqi; Hu, Minyu; Liu, Shaohua; Wang, Yuxiang; Gao, Qile; Guo, Chaofeng

    2018-01-01

    The proliferative rate of chondrocytes affects bone elongation. Chondrocyte hypertrophy is required for endochondral bone formation as chondrocytes secrete factors required for osteoblast differentiation and maturation. Previous studies have demonstrated that the Indian hedgehog (Ihh) signaling pathway is a key regulator of skeletal development and homeostasis. The aim of the present study was to investigate the function of Ihh in chondrocyte proliferation and differentiation, as well as the underlying mechanisms. Ihh was knocked down in mouse chondrocyte cells using short hairpin RNA. Chondrocyte apoptosis and cell cycle arrest were assessed using flow cytometry and the results indicated that knockdown of Ihh significantly inhibited cell growth (PIhh also resulted in cell cycle arrest at G1 to S phase in chondrocytes. It was also observed that knockdown of Ihh decreased alkaline phosphatase activity and mineral deposition of chondrocytes. The inhibitory roles of Ihh downregulation on chondrocyte growth and differentiation may be associated with the transforming growth factor-β/mothers against decapentaplegic and osteoprotegerin/receptor activator of nuclear factor κB ligand signaling pathway. The results of the present study suggest that chondrocyte-derived Ihh is essential for maintaining bone growth plates and that manipulation of Ihh expression or its signaling components may be a novel therapeutic technique for the treatment of skeletal diseases, including achondroplasia.

  1. RHEB: a potential regulator of chondrocyte phenotype for cartilage tissue regeneration.

    Science.gov (United States)

    Ashraf, S; Ahn, J; Cha, B-H; Kim, J-S; Han, I; Park, H; Lee, S-H

    2017-09-01

    As articular cartilage has a limited ability to self-repair, successful cartilage regeneration requires clinical-grade chondrocytes with innate characteristics. However, cartilage regeneration via chondrocyte transplantation is challenging, because chondrocytes lose their innate characteristics during in vitro expansion. Here, we investigated the mechanistic underpinning of the gene Ras homologue enriched in brain (RHEB) in the control of senescence and dedifferentiation through the modulation of oxidative stress in chondrocytes, a hallmark of osteoarthritis. Serial expansion of human chondrocytes led to senescence, dedifferentiation and oxidative stress. RHEB maintained the innate characteristics of chondrocytes by regulating senescence, dedifferentiation and oxidative stress, leading to the upregulation of COL2 expression via SOX9 and the downregulation of p27 expression via MCL1. RHEB also decreased the expression of COL10. RHEB knockdown mimics decreased the expression of SOX9, COL2 and MCL1, while abrogating the suppressive function of RHEB on p27 and COL10 in chondrocytes. RHEB-overexpressing chondrocytes successfully formed cartilage tissue in vitro as well as in vivo, with increased expression of GAG matrix and chondrogenic markers. RHEB induces a distinct gene expression signature that maintained the innate chondrogenic properties over a long period. Therefore, RHEB expression represents a potentially useful mechanism in terms of cartilage tissue regeneration from chondrocytes, by which chondrocyte phenotypic and molecular characteristics can be retained through the modulation of senescence, dedifferentiation and oxidative stress. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  2. Role of interleukin-1 in antigen presentation by normal articular chondrocytes

    International Nuclear Information System (INIS)

    Tiku, M.L.; Liu, S.; Tiku, K.

    1986-01-01

    Recently the authors have described that normal articular chondrocytes of rabbits present antigen to immune T cells. In the present study the authors investigated the role of interleukin-1 (IL-1) on antigen presentation by chondrocytes. For these experiments the antigen pulsed chondroyctes were either untreated or fixed with paraformaldehyde and then co-cultured with immune T cells. T cell proliferation was measured by 3 H-thymidine incorporation. Pulsed non-fixed chondrocytes presented antigen, as expected, but pulsed and fixed cells failed to present antigen to T cells. The 3 H-TdR incorporation was partially restored by addition of purified human IL-1. Next, IL-1 activity was measured in primary chondrocyte culture supernatants stimulated with or without lipopolysaccharide (LPS) in comitogen thymocyte assay. No activity was detected in chondrocyte supernatants. Propagated chondrocyte culture supernatants also lacked IL-1 activity when stimulated with LPS in the presence of increasing concentration of indomethacin. On the other hand the authors observed that chondrocyte culture supernatants in a dose dependent manner inhibited human IL-1 induced 3 H-TdR incorporation of murine thymocytes. This suggested that these cells may produce an inhibitor of IL-1 and IL-1 production by chondrocytes may be essential for T cell proliferation by these cells. Inability to detect IL-1 in chondrocyte supernatants may be due to the presence of an inhibitor to IL-1. These findings may help in elucidating the immunological mechanisms in situations where chondrocytes and T cell interact, such as in arthritis

  3. Indian hedgehog signaling triggers Nkx3.2 protein degradation during chondrocyte maturation

    Science.gov (United States)

    Choi, Seung-Won; Jeong, Da-Un; Kim, Jeong-Ah; Lee, Boyoung; Joeng, Kyu Sang; Long, Fanxin; Kim, Dae-Won

    2015-01-01

    The Indian hedgehog (Ihh) pathway plays an essential role in facilitating chondrocyte hypertrophy and bone formation during skeletal development. Nkx3.2 is initially induced in chondrocyte precursor cells, maintained in early-stage chondrocytes, and down-regulated in terminal-stage chondrocytes. Consistent with these expression patterns, Nkx3.2 has been shown to enhance chondrocyte differentiation and cell survival, while inhibiting chondrocyte hypertrophy and apoptosis. Thus, in this work, we investigate whether Nkx3.2, an early stage chondrogenic factor, can be regulated by Ihh, a key regulator for chondrocyte hypertrophy. Here, we show that Ihh signaling can induce proteasomal degradation of Nkx3.2. In addition, we found that Ihh can suppress levels of Lrp (Wnt co-receptor) and Sfrp (Wnt antagonist) expression, which, in turn, may selectively enhance Lrp-independent non-canonical Wnt pathways in chondrocyte. In agreement with these findings, Ihh-induced Nkx3.2 degradation requires Wnt5a, which is capable of triggering Nkx3.2 degradation. Finally, we found that Nkx3.2 protein levels in chondrocytes are remarkably elevated in mice defective in Ihh signaling by deletion of either Ihh or Smoothened. Thus, these results suggest that Ihh/Wnt5a signaling may play a role in negative regulation of Nkx3.2 for appropriate progression of chondrocyte hypertrophy during chondrogenesis. PMID:22507129

  4. Conditional expression of constitutively active estrogen receptor {alpha} in chondrocytes impairs longitudinal bone growth in mice

    Energy Technology Data Exchange (ETDEWEB)

    Ikeda, Kazuhiro [Division of Gene Regulation and Signal Transduction, Research Center for Genomic Medicine, Saitama Medical University, Saitama (Japan); Tsukui, Tohru [Experimental Animal Laboratory, Research Center for Genomic Medicine, Saitama Medical University, Saitama (Japan); Imazawa, Yukiko; Horie-Inoue, Kuniko [Division of Gene Regulation and Signal Transduction, Research Center for Genomic Medicine, Saitama Medical University, Saitama (Japan); Inoue, Satoshi, E-mail: INOUE-GER@h.u-tokyo.ac.jp [Division of Gene Regulation and Signal Transduction, Research Center for Genomic Medicine, Saitama Medical University, Saitama (Japan); Department of Geriatric Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo (Japan); Department of Anti-Aging Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo (Japan)

    2012-09-07

    Highlights: Black-Right-Pointing-Pointer Conditional transgenic mice expressing constitutively active estrogen receptor {alpha} (caER{alpha}) in chondrocytes were developed. Black-Right-Pointing-Pointer Expression of caER{alpha} in chondrocytes impaired longitudinal bone growth in mice. Black-Right-Pointing-Pointer caER{alpha} affects chondrocyte proliferation and differentiation. Black-Right-Pointing-Pointer This mouse model is useful for understanding the physiological role of ER{alpha}in vivo. -- Abstract: Estrogen plays important roles in the regulation of chondrocyte proliferation and differentiation, which are essential steps for longitudinal bone growth; however, the mechanisms of estrogen action on chondrocytes have not been fully elucidated. In the present study, we generated conditional transgenic mice, designated as caER{alpha}{sup ColII}, expressing constitutively active mutant estrogen receptor (ER) {alpha} in chondrocytes, using the chondrocyte-specific type II collagen promoter-driven Cre transgenic mice. caER{alpha}{sup ColII} mice showed retardation in longitudinal growth, with short bone lengths. BrdU labeling showed reduced proliferation of hypertrophic chondrocytes in the proliferating layer of the growth plate of tibia in caER{alpha}{sup ColII} mice. In situ hybridization analysis of type X collagen revealed that the maturation of hypertrophic chondrocytes was impaired in caER{alpha}{sup ColII} mice. These results suggest that ER{alpha} is a critical regulator of chondrocyte proliferation and maturation during skeletal development, mediating longitudinal bone growth in vivo.

  5. Conditional expression of constitutively active estrogen receptor α in chondrocytes impairs longitudinal bone growth in mice

    International Nuclear Information System (INIS)

    Ikeda, Kazuhiro; Tsukui, Tohru; Imazawa, Yukiko; Horie-Inoue, Kuniko; Inoue, Satoshi

    2012-01-01

    Highlights: ► Conditional transgenic mice expressing constitutively active estrogen receptor α (caERα) in chondrocytes were developed. ► Expression of caERα in chondrocytes impaired longitudinal bone growth in mice. ► caERα affects chondrocyte proliferation and differentiation. ► This mouse model is useful for understanding the physiological role of ERαin vivo. -- Abstract: Estrogen plays important roles in the regulation of chondrocyte proliferation and differentiation, which are essential steps for longitudinal bone growth; however, the mechanisms of estrogen action on chondrocytes have not been fully elucidated. In the present study, we generated conditional transgenic mice, designated as caERα ColII , expressing constitutively active mutant estrogen receptor (ER) α in chondrocytes, using the chondrocyte-specific type II collagen promoter-driven Cre transgenic mice. caERα ColII mice showed retardation in longitudinal growth, with short bone lengths. BrdU labeling showed reduced proliferation of hypertrophic chondrocytes in the proliferating layer of the growth plate of tibia in caERα ColII mice. In situ hybridization analysis of type X collagen revealed that the maturation of hypertrophic chondrocytes was impaired in caERα ColII mice. These results suggest that ERα is a critical regulator of chondrocyte proliferation and maturation during skeletal development, mediating longitudinal bone growth in vivo.

  6. Clinical and immune characteristics of men, who worked within 30-km zone near Chernobyl four years after the accident

    International Nuclear Information System (INIS)

    Oradovskaya, I.V.; Ruzybakiev, R.M.; Prokopenko, V.D.

    1994-01-01

    Clinical examination with immune status assessment of more than 300 healthy persons, who worked in 30-km Chernobyl NPP zone four years after the accident was carried out. The clinical characters of immune deficiency were detected in 6.67 % of examined persons and appeared as repeated respiratory diseases. Significant increase in mean level of leukocytes and lymphocytes was revealed. Activation of T-helper link of cell immunity was determined by increase of CD 4 + cells in 35.2 % of examined persons, that were lower than in people who took part in Chernobyl accident amelioration in 1986. Decrease in content of T-cells and B-cells was observed in 1/4 and 3/4 of persons correspondingly by rosette assays. No changes in serum immunoglobulins level was observed

  7. Mammary tumors and serum hormones in the bitch treated with medroxyprogesterone acetate or progesterone for four years

    Energy Technology Data Exchange (ETDEWEB)

    Frank, D.W.; Kirton, K.T.; Murchison, T.E.; Quinlan, W.J.; Coleman, M.E.; Gilbertson, T.J.; Feenstra, E.S.; Kimball, F.A.

    1978-01-01

    After four years of a long term contraceptive steroid safety study, the incidence and the histologic type of mammary dysplasia produced is similar in beagles treated with medroxyprogesterone acetate (medroxyprogesterone) or progesterone. Serum insulin, thyroid stimulating hormone (TSH), triiodothyronine, growth hormone, prolactin, 17..beta..-estradiol, progesterone, and cortisol were determined by radioimmunoassay on samples collected after 45 months of treatment. Serum growth hormone and insulin concentrations were elevated in a dose related manner in both treatment groups. Triiodothyronine, cortisol, and estradiol-17..beta.. (medroxyprogesterone only) were lowered. TSH and prolactin concentrations were not changed. Pituitary--gonadal hormone interaction in the pathogenesis of mammary neoplasia of the dog is discussed. Prolonged treatment of the beagle with massive doses of progesterone or medroxyprogesterone results in a dose related incidence of mammary modules.

  8. The national neonatal screening programme for congenital toxoplasmosis in Denmark: results from the initial four years, 1999-2002

    DEFF Research Database (Denmark)

    Schmidt, Dorte Remmer; Høgh, Birthe; Andersen, O

    2006-01-01

    AIMS: To describe the outcome of four years' nationwide neonatal screening for congenital toxoplasmosis in liveborn newborns. METHODS: Congenital toxoplasmosis was diagnosed if specific Toxoplasma gondii IgM antibodies were detected in eluate from the PKU Guthrie filter paper card from a child....... Infants diagnosed with congenital toxoplasmosis were examined for intracranial and retinal lesions and treated for three months with sulphadiazine, pyrimethamine, and folinic acid continuously. RESULTS: Eluates from PKU-cards from 262 912 newborns were analysed. The birth prevalence of congenital...... toxoplasma infection was 2.1 per 10 000 liveborns. Congenital toxoplasmosis was suspected in 96 infants and confirmed in 55. Forty seven children were examined for intracranial and retinal lesions soon after birth; 12 had clinical signs at this first examination. Of these, 5 had intracranial calcifications...

  9. Correlates of light and moderate-to-vigorous objectively measured physical activity in four-year-old children.

    Science.gov (United States)

    van Sluijs, Esther M F; McMinn, Alison M; Inskip, Hazel M; Ekelund, Ulf; Godfrey, Keith M; Harvey, Nicholas C; Griffin, Simon J

    2013-01-01

    Correlates of physical activity (PA) are hypothesized to be context and behaviour specific, but there is limited evidence of this in young children. The aim of the current study is to investigate associations between personal, social and environmental factors and objectively measured light and moderate-to-vigorous PA (LPA and MVPA, respectively) in four-year-old children. Cross-sectional data were used from the Southampton Women's Survey, a UK population-based longitudinal study. Four-year old children (n = 487, 47.0% male) had valid PA data assessed using accelerometry (Actiheart) and exposure data collected with a validated maternal questionnaire (including data on child personality, family demographics, maternal behaviour, rules and restrictions, and perceived local environment). Linear regression modelling was used to analyse associations with LPA and MVPA separately, interactions with sex were explored. LPA minutes were greater in children whose mothers reported more PA (vs. inactive: regression coefficient±standard error: 6.70±2.94 minutes), and without other children in the neighbourhood to play with (-6.33±2.44). MVPA minutes were greater in children with older siblings (vs. none: 5.81±2.80) and those whose mothers used active transport for short trips (vs. inactive: 6.24±2.95). Children accumulated more MVPA in spring (vs. winter: 9.50±4.03) and, in boys only, less MVPA with availability of other children in the neighbourhood (-3.98±1.70). Young children's LPA and MVPA have differing associations with a number of social and environmental variables. Interventions targeting PA promotion in young children outside of formal care settings should consider including intensity specific factors.

  10. Correlates of light and moderate-to-vigorous objectively measured physical activity in four-year-old children.

    Directory of Open Access Journals (Sweden)

    Esther M F van Sluijs

    Full Text Available BACKGROUND: Correlates of physical activity (PA are hypothesized to be context and behaviour specific, but there is limited evidence of this in young children. The aim of the current study is to investigate associations between personal, social and environmental factors and objectively measured light and moderate-to-vigorous PA (LPA and MVPA, respectively in four-year-old children. METHODS: Cross-sectional data were used from the Southampton Women's Survey, a UK population-based longitudinal study. Four-year old children (n = 487, 47.0% male had valid PA data assessed using accelerometry (Actiheart and exposure data collected with a validated maternal questionnaire (including data on child personality, family demographics, maternal behaviour, rules and restrictions, and perceived local environment. Linear regression modelling was used to analyse associations with LPA and MVPA separately, interactions with sex were explored. RESULTS: LPA minutes were greater in children whose mothers reported more PA (vs. inactive: regression coefficient±standard error: 6.70±2.94 minutes, and without other children in the neighbourhood to play with (-6.33±2.44. MVPA minutes were greater in children with older siblings (vs. none: 5.81±2.80 and those whose mothers used active transport for short trips (vs. inactive: 6.24±2.95. Children accumulated more MVPA in spring (vs. winter: 9.50±4.03 and, in boys only, less MVPA with availability of other children in the neighbourhood (-3.98±1.70. DISCUSSION: Young children's LPA and MVPA have differing associations with a number of social and environmental variables. Interventions targeting PA promotion in young children outside of formal care settings should consider including intensity specific factors.

  11. MR imaging of autologous chondrocyte implantation of the knee

    Energy Technology Data Exchange (ETDEWEB)

    James, S.L.J.; Connell, D.A.; Saifuddin, A.; Skinner, J.A.; Briggs, T.W.R. [RNOH Stanmore, Department of Radiology, Stanmore, Middlesex (United Kingdom)

    2006-05-15

    Autologous chondrocyte implantation (ACI) is a surgical technique that is increasingly being used in the treatment of full-thickness defects of articular cartilage in the knee. It involves the arthroscopic harvesting and in vitro culture of chondrocytes that are subsequently implanted into a previously identified chondral defect. The aim is to produce a repair tissue that closely resembles hyaline articular cartilage that gradually becomes incorporated, restoring joint congruity. Over the long term, it is hoped that this will prevent the progression of full-thickness articular cartilage defects to osteoarthritis. This article reviews the indications and operative procedure performed in ACI. Magnetic resonance imaging (MRI) sequences that provide optimal visualization of articular cartilage in the post-operative period are discussed. Normal appearances of ACI on MRI are presented along with common complications that are encountered with this technique. (orig.)

  12. Can microcarrier-expanded chondrocytes synthesize cartilaginous tissue in vitro?

    Science.gov (United States)

    Surrao, Denver C; Khan, Aasma A; McGregor, Aaron J; Amsden, Brian G; Waldman, Stephen D

    2011-08-01

    Tissue engineering is a promising approach for articular cartilage repair; however, it is challenging to produce adequate amounts of tissue in vitro from the limited number of cells that can be extracted from an individual. Relatively few cell expansion methods exist without the problems of de-differentiation and/or loss of potency. Recently, however, several studies have noted the benefits of three-dimensional (3D) over monolayer expansion, but the ability of 3D expanded chondrocytes to synthesize cartilaginous tissue constructs has not been demonstrated. Thus, the purpose of this study was to compare the properties of engineered cartilage constructs from expanded cells (monolayer and 3D microcarriers) to those developed from primary chondrocytes. Isolated bovine chondrocytes were grown for 3 weeks in either monolayer (T-Flasks) or 3D microcarrier (Cytodex 3) expansion culture. Expanded and isolated primary cells were then seeded in high density culture on Millicell™ filters for 4 weeks to evaluate the ability to synthesize cartilaginous tissue. While microcarrier expansion was twice as effective as monolayer expansion (microcarrier: 110-fold increase, monolayer: 52-fold increase), the expanded cells (monolayer and 3D microcarrier) were not effectively able to synthesize cartilaginous tissue in vitro. Tissues developed from primary cells were substantially thicker and accumulated significantly more extracellular matrix (proteoglycan content: 156%-292% increase; collagen content: 70%-191% increase). These results were attributed to phenotypic changes experienced during the expansion phase. Monolayer expanded chondrocytes lost their native morphology within 1 week, whereas microcarrier-expanded cells were spreading by 3 weeks of expansion. While the use of 3D microcarriers can lead to large cellular yields, preservation of chondrogenic phenotype during expansion is required in order to synthesize cartilaginous tissue.

  13. Statins do not inhibit the FGFR signaling in chondrocytes

    Czech Academy of Sciences Publication Activity Database

    Fafílek, B.; Hampl, Marek; Říčánková, N.; Veselá, Iva; Bálek, L.; Kunová Bosáková, M.; Gudernová, I.; Vařecha, M.; Buchtová, Marcela; Krejčí, P.

    2017-01-01

    Roč. 25, č. 9 (2017), s. 1522-1530 ISSN 1063-4584 R&D Projects: GA ČR(CZ) GA14-31540S Grant - others:GA MŠk(CZ) LH12004 Institutional support: RVO:67985904 Keywords : statins * FGF signaling * chondrocytes Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Developmental biology Impact factor: 4.742, year: 2016

  14. The effect of cartilage and bone density of mushroom-shaped, photooxidized, osteochondral transplants: an experimental study on graft performance in sheep using transplants originating from different species

    Directory of Open Access Journals (Sweden)

    Hilbe Monika

    2005-12-01

    Full Text Available Abstract Background Differences in overall performance of osteochondral photooxidized grafts were studied in accordance of their species origin and a new, more rigorous cleansing procedure using alcohol during preparation. Methods Photooxidized mushroom-shaped grafts of bovine, ovine, human and equine origin were implanted in the femoral condyles of 32 sheep (condyles: n = 64. No viable chondrocytes were present at the time of implantation. Grafts were evaluated at 6 months using plastic embedded sections of non-decalcified bone and cartilage specimens. Graft incorporation, the formation of cyst-like lesions at the base of the cartilage junction as well as cartilage morphology was studied qualitatively, semi-quantitatively using a score system and quantitatively by performing histomorphometrical measurements of percentage of bone and fibrous tissue of the original defects. For statistical analysis a factorial analysis of variance (ANOVA- test was applied. Results Differences of graft performance were found according to species origin and cleansing process during graft preparation. According to the score system cartilage surface integrity was best for equine grafts, as well as dislocation or mechanical stability. The equine grafts showed the highest percentage for bone and lowest for fibrous tissue, resp. cystic lesions. The new, more rigorous cleansing process decreased cartilage persistence and overall graft performance. Conclusion Performance of grafts from equine origin was better compared to bovine, ovine and human grafts. The exact reason for this difference was not proven in the current study, but could be related to differences in density of cartilage and subchondral bone between species.

  15. Regulation of collagenase inhibitor production in chondrosarcoma chondrocytes

    International Nuclear Information System (INIS)

    Harper, J.; Harper, E.

    1987-01-01

    Swarm rat chondrosarcoma chondrocytes produce an inhibitor of collagenase. This inhibitor is similar to those isolated from normal cartilage tissues. These cells will synthesize proteins in the absence of serum. Since serum contains inhibitors of collagenase, it is necessary to culture cells without serum in order to obtain accurate measurements of enzyme and inhibitor levels. They examined the effect of insulin on inhibitor secretion by cultures of Swarm rat chondrosarcoma chondrocytes. They observed a 2.5 to 3.5 fold stimulation of inhibitory activity in the presence of as little as 10 ng/ml insulin as compared to controls in serum free Dulbecco's modified Eagle's medium supplemented with 4.5 g/l glucose. The units of inhibitor were determined over a 7 day culture period. Medium was harvested daily and assayed for collagenase activity and for inhibition of a known collagenase from rabbit skin or human skin, using the 14 C-glycine peptide release assay. The amount of inhibitor obtained from days 2 through 7 were: 1.4 unit (control), 3.8 units (10 ng/ml insulin), 5.2 units (1 μg/ml insulin). The addition of 1 mM dibutyryl cyclic AMP to these chondrocytes in the presence of 1 μg/ml insulin caused a decrease in the level of inhibitor, suggesting that a dephosphorylation event may be necessary for this stimulation by insulin to occur

  16. Hydrostatic Pressure Influences HIF-2 Alpha Expression in Chondrocytes

    Directory of Open Access Journals (Sweden)

    Hiroaki Inoue

    2015-01-01

    Full Text Available Hypoxia-inducible factor (HIF-2α is considered to play a major role in the progression of osteoarthritis. Recently, it was reported that pressure amplitude influences HIF-2α expression in murine endothelial cells. We examined whether hydrostatic pressure is involved in expression of HIF-2α in articular chondrocytes. Chondrocytes were cultured and stimulated by inflammation or hydrostatic pressure of 0, 5, 10, or 50 MPa. After stimulation, heat shock protein (HSP 70, HIF-2α, nuclear factor kappa B (NF-κB, matrix metalloproteinase (MMP-13, MMP-3, and vascular endothelial growth factor (VEGF gene expression were evaluated. The levels of all gene expression were increased by inflammatory stress. When chondrocytes were exposed to a hydrostatic pressure of 5 MPa, HIF-2α, MMP-13, and MMP-3 gene expression increased significantly although those of HSP70 and NF-κB were not significantly different from the control group. In contrast, HIF-2α gene expression did not increase under a hydrostatic pressure of 50 MPa although HSP70 and NF-κB expression increased significantly compared to control. We considered that hydrostatic pressure of 5 MPa could regulate HIF-2α independent of NF-κB, because the level of HIF-2α gene expression increased significantly without upregulation of NF-κB expression at 5 MPa. Hydrostatic pressure may influence cartilage degeneration, inducing MMP-13 and MMP-3 expression through HIF-2α.

  17. Hydrostatic pressure influences HIF-2 alpha expression in chondrocytes.

    Science.gov (United States)

    Inoue, Hiroaki; Arai, Yuji; Kishida, Tsunao; Terauchi, Ryu; Honjo, Kuniaki; Nakagawa, Shuji; Tsuchida, Shinji; Matsuki, Tomohiro; Ueshima, Keiichirou; Fujiwara, Hiroyoshi; Mazda, Osam; Kubo, Toshikazu

    2015-01-05

    Hypoxia-inducible factor (HIF)-2α is considered to play a major role in the progression of osteoarthritis. Recently, it was reported that pressure amplitude influences HIF-2α expression in murine endothelial cells. We examined whether hydrostatic pressure is involved in expression of HIF-2α in articular chondrocytes. Chondrocytes were cultured and stimulated by inflammation or hydrostatic pressure of 0, 5, 10, or 50 MPa. After stimulation, heat shock protein (HSP) 70, HIF-2α, nuclear factor kappa B (NF-κB), matrix metalloproteinase (MMP)-13, MMP-3, and vascular endothelial growth factor (VEGF) gene expression were evaluated. The levels of all gene expression were increased by inflammatory stress. When chondrocytes were exposed to a hydrostatic pressure of 5 MPa, HIF-2α, MMP-13, and MMP-3 gene expression increased significantly although those of HSP70 and NF-κB were not significantly different from the control group. In contrast, HIF-2α gene expression did not increase under a hydrostatic pressure of 50 MPa although HSP70 and NF-κB expression increased significantly compared to control. We considered that hydrostatic pressure of 5 MPa could regulate HIF-2α independent of NF-κB, because the level of HIF-2α gene expression increased significantly without upregulation of NF-κB expression at 5 MPa. Hydrostatic pressure may influence cartilage degeneration, inducing MMP-13 and MMP-3 expression through HIF-2α.

  18. Doublecortin May Play a Role in Defining Chondrocyte Phenotype

    Directory of Open Access Journals (Sweden)

    Dongxia Ge

    2014-04-01

    Full Text Available Embryonic development of articular cartilage has not been well understood and the role of doublecortin (DCX in determination of chondrocyte phenotype is unknown. Here, we use a DCX promoter-driven eGFP reporter mouse model to study the dynamic gene expression profiles in mouse embryonic handplates at E12.5 to E13.5 when the condensed mesenchymal cells differentiate into either endochondral chondrocytes or joint interzone cells. Illumina microarray analysis identified a variety of genes that were expressed differentially in the different regions of mouse handplate. The unique expression patterns of many genes were revealed. Cytl1 and 3110032G18RIK were highly expressed in the proximal region of E12.5 handplate and the carpal region of E13.5 handplate, whereas Olfr538, Kctd15, and Cited1 were highly expressed in the distal region of E12.5 and the metacarpal region of E13.5 handplates. There was an increasing gradient of Hrc expression in the proximal to distal direction in E13.5 handplate. Furthermore, when human DCX protein was expressed in human adipose stem cells, collagen II was decreased while aggrecan, matrilin 2, and GDF5 were increased during the 14-day pellet culture. These findings suggest that DCX may play a role in defining chondrocyte phenotype.

  19. Exploring the experiences of female students in introductory project-based engineering courses at two- and four-year institutions

    Science.gov (United States)

    Swan, Amy K.

    2011-12-01

    This qualitative study explored the experiential and contextual factors that shaped female students' pathways into introductory project-based engineering classes at two community colleges and one four-year institution, as well as female students' experiences within and outside of these classes. The study was framed by Social Cognitive Career Theory (SCCT) (Lent, Brown & Hackett, 1996) and Bronfenbrenner's (1979) ecological systems theory. Findings were based on analyses of data gathered through multiple methods: observations; individual interviews with female students; focus group interviews with project teams; and document collection. The findings of this study revealed that while positive experiences with math or science were a likely pre-cursor to engineering interest, experiential learning appeared to be a more powerful force in fostering students' engineering interest. Specifically, participants developed an interest in engineering through academic, professional, and extracurricular engineering- and design-related activities that familiarized them with the tasks and skills involved in engineering work and helped them develop a sense of selfefficacy with regard to this work. Interest and self-efficacy, in turn, played a role in students' postsecondary educational decision-making processes, as did contextual factors including families and finances. This study's findings also showed that participants' project teams were a critically important microsystem within participants' ecological environments. Within this sometimes "chilly" microsystem, female students negotiated intrateam processes, which were in some cases affected by gender norms. Intrateam processes that influenced female students' project-based learning experiences included: interpersonal dynamics; leadership; and division of labor. This study also identified several ways in which the lived experiences of participants at the community colleges were different from, or similar to, those of participants

  20. IGF1 regulates RUNX1 expression via IRS1/2: Implications for antler chondrocyte differentiation

    OpenAIRE

    Yang, Zhan-Qing; Zhang, Hong-Liang; Duan, Cui-Cui; Geng, Shuang; Wang, Kai; Yu, Hai-Fan; Yue, Zhan-Peng; Guo, Bin

    2017-01-01

    Although IGF1 is important for the proliferation and differentiation of chondrocytes, its underlying molecular mechanism is still unknown. Here we addressed the physiologic function of IGF1 in antler cartilage and explored the interplay of IGF1, IRS1/2 and RUNX1 in chondrocyte differentiation. The results showed that IGF1 was highly expressed in antler chondrocytes. Exogenous rIGF1 could increase the proliferation of chondrocytes and cell proportion in the S phase, whereas IGF1R inhibitor PQ4...

  1. Training Intensity Distribution Over a Four-Year Cycle in Olympic Champion Rowers: Different Roads Lead to Rio.

    Science.gov (United States)

    Plews, Daniel J; Laursen, Paul B

    2017-09-27

    The purpose of this study was to compare the training intensity distribution (TID) of the undefeated world champion male rowing New Zealand (kiwi) pair over a four-year Olympic cycle, across training phases, training years, and between individuals. Training data, including heart rate and boat speed, were recorded in the athletes rowing in the same boat between March 2013 and August 2016, ending with the Rio Olympics final. Progressive exercise tests assessed first (LT 1 ) and second (LT 2 ) lactate thresholds and associated heart rates, to determine the percentage of training performed below, between and above these demarcation points. Training an average of only 12-15 h/wk throughout the Olympic cycle, the mean percent distribution of time (±SD) at each training intensity was 80.4 ± 5.5% LT 2 for Rower A and 67.3 ± 9.0% LT 2 for Rower B. Across the years 2014-2016, Rower A performed most likely more training training between LT 1 -LT 2 . Training appeared to become more polarised, with greater amounts of time spent training duration (R=0.38-0.43). Two of the world's best rowers, rowing together in the same boat with an undefeated record across an Olympic cycle, travelled markedly different "roads to Rio" within the context of their TID, with one rower displaying a polarised model of TID, and the other pyramidal. However, TID trended towards becoming more polarised in both rowers with increased training duration.

  2. New CLGF Four-Year Grant to Help Local Government Service Delivery and Boost CLGF’s Research Capacity

    Directory of Open Access Journals (Sweden)

    Lucy Slack

    2012-12-01

    Full Text Available The UK Department for International Development (UK AID has agreed £4.5 million funding for a four-year CLGF programme to improve governance and service delivery at local level in several areas of the Commonwealth including Africa and Asia from 2012-16. It will also help to support national policy frameworks for local government service delivery, and increase engagement of local government in regional policy planning and implementation. CLGF will continue to work with its members, UN partners and others to mobilise more resources towards the support of local government in the Commonwealth. The new programme will focus on local government pilot projects in LED, supporting ministries and local government associations in strengthening their national policy making for local government, and establish regional forums to enable local government to engage in and influence regional policy making to reflect the needs and priorities of local government. It will also boost CLGF’s research capacity with targeted research to strengthen CLGF’s policy making and advocacy, including more sustained engagement in international policy debates on key issues affecting local government, such as climate change.

  3. Integrating critical thinking and evidence-based dentistry across a four-year dental curriculum: a model for independent learning.

    Science.gov (United States)

    Marshall, Teresa A; Straub-Morarend, Cheryl L; Handoo, Nidhi; Solow, Catherine M; Cunningham-Ford, Marsha A; Finkelstein, Michael W

    2014-03-01

    Introducing critical thinking and evidence-based dentistry (EBD) content into an established dental curriculum can be a difficult and challenging process. Over the past three years, the University of Iowa College of Dentistry has developed and implemented a progressive four-year integrated critical thinking and EBD curriculum. The objective of this article is to describe the development and implementation process to make it available as a model for other dental schools contemplating introduction of critical thinking and EBD into their curricula. The newly designed curriculum built upon an existing problem-based learning foundation, which introduces critical thinking and the scientific literature in the D1 year, in order to expose students to the rationale and resources for practicing EBD in the D2 and D3 years and provide opportunities to practice critical thinking and apply the EBD five-step process in the D2, D3, and D4 years. All curricular content is online, and D3 and D4 EBD activities are integrated within existing clinical responsibilities. The curricular content, student resources, and student activities are described.

  4. Four years of experience with the use of calculated isotopic correlations in establishing input balances at the La Hague plant

    International Nuclear Information System (INIS)

    Aries, M.; Patigny, P.; Bouchard, J.; Giacometti, A.; Girieud, R.

    1983-01-01

    For more than four years the La Hague reprocessing plant has been using calculated isotopic correlations to establish and check its input balances. The masses of uranium and plutonium entering the plant are determined by the gravimetric balance method, which utilizes the burnup obtained by calculated isotopic correlation as well as the Pu/U ratio measured at the dissolver after cross-checking with the values obtained by correlation. Further, a verification of all the parameters needed to establish these balances - whether physical or chemical in origin - is carried out systematically by means of internal coherence constants which make it possible to detect any anomalies in the dissolution data. The calculated isotopic correlations were evaluated when the analyses of numerous representative samples of irradiated fuel and experimental results of separated isotopic irradiation in water reactor spectra had been interpreted. The accuracy achieved was improved by allowing in the neutron calculations for effects inherent in the first reactor core and by selecting a set of calculation functions which attenuates (by compensation effects) the various perturbations in the irradiation history. The results obtained at La Hague with calculated isotopic correlations on nearly 600 t of reprocessed UO 2 , because of their large number and above all their high quality, suggest that it be proposed extending the method to other reprocessing plants. This could be done by the operator himself or by national or international control bodies within the framework of a safeguards arrangement. (author)

  5. [Intravenous thrombolysis for acute ischemic stroke. A four years’ experience in a Chilean public hospital].

    Science.gov (United States)

    Soto V, Álvaro; Morales I, Gladys; Grandjean B, Marcela; Pollak W, Débora; Del Castillo C, Carolina; García F, Pía; Von Johnn A, Alexis; Riquelme G, Alfonso

    2017-04-01

    Intravenous thrombolysis (IVT) with alteplase (tissue plasminogen activator) is the standard pharmacological treatment in acute ischemic stroke (AIS), reducing disability in patients. To report the results a thrombolysis protocol during four years in a regional public hospital. Data from 106 consecutive patients aged 68 ± 13 years (57% men) who were treated with IVT, from May 2012 until April 2016, was analyzed. The median door-to-needle time was 80 minutes (interquartile range = 57-113). The median National Institute of Health Stroke Scale (NIHSS) scores on admission and at discharge were was 11.5 and 5 points respectively. At discharge, 27% of hospitalized patients had a favorable outcome (n = 99), defined as having 0 to 1 points in the modified Rankin scale. Symptomatic intracerebral hemorrhage and mortality rates were 5.7 and 13.1%, respectively. The thrombolysis rate rose from 0.7% in 2012 to 6% in 2016. The implementation of 24/7 neurology shifts in the Emergency Department allowed us to increase the amount and quality of IVT in our hospital, as measured by the rate of thrombolysis and by process indicators such as door-to-needle time.

  6. Bacterial Pathogens and Antimicrobial Resistance Patterns in Pediatric Urinary Tract Infections: A Four-Year Surveillance Study (2009–2012

    Directory of Open Access Journals (Sweden)

    Seyed Reza Mirsoleymani

    2014-01-01

    Full Text Available The aims of this study were to assess the common bacterial microorganisms causing UTI and their antimicrobial resistance patterns in Bandar Abbas (Southern Iran during a four-year period. In this retrospective study, samples with a colony count of ≥105 CFU/mL bacteria were considered positive; for these samples, the bacteria were identified, and the profile of antibiotic susceptibility was characterized. From the 19223 samples analyzed, 1513 (7.87% were positive for bacterial infection. UTI was more frequent in male (54.9%. E. coli was reported the most common etiological agent of UTI (65.2%, followed by Klebsiella spp. (26%, Pseudomonas aeruginosa (3.6%, and Staphylococcus coagulase positive (3.7%. Results of antimicrobial susceptibility analysis for E. coli to commonly used antibiotics are as follows: Amikacin (79.7%, Ofloxacin (78.3%, Gentamicin (71.6%, Ceftriaxone (41.8, Cefotaxime (41.4%, and Cefixime (27.8%. Empirical antibiotic selection should be based on awareness of the local prevalence of bacterial organisms and antibiotic sensitivities rather than on universal or even national guidelines. In this study, Amikacin and Gentamicin were shown to be the most appropriate antibiotics for empiric therapy of pyelonephritis, but empirical therapy should only be done by specialist physicians in cases where it is necessary while considering sex and age of children.

  7. Four year experience of sarcoma of soft tissues and bones in a tertiary care hospital and review of literature

    Directory of Open Access Journals (Sweden)

    Ansari Tayyaba Z

    2011-05-01

    Full Text Available Abstract Background Sarcoma encompasses an uncommon group of cancer and the data is insufficient from Pakistan. We report our four years experience of Sarcoma of soft tissues and bones. Methods This cross sectional study was carried out at Aga Khan University Hospital from 2004 to 2008. The patients were divided into two groups from the outset i.e. initially diagnosed and relapsed group and separate sub group analysis was conducted. Results Out of 93 newly diagnosed patients, 58 belonged to bone sarcoma and 35 to soft tissue sarcoma group. While for relapsed patients, 5 had soft tissue sarcoma and 9 had bone sarcoma. Mean age was 32.5 years. At presentation, approximately two third patients had localised disease while remaining one third had metastatic disease. The Kaplan Meier estimate of median recurrence free survival was 25 months, 35 months, and 44 months for Osteogenic sarcoma, Ewing's sarcoma and Chondrosarcoma respectively. For Leiomyosarcoma and Synovial sarcoma, it was 20 and 19 months respectively. The grade of the tumour (p = 0.02 and surgical margin status (p = 0.001 were statistically significant for determination of relapse of disease. Conclusion The median recurrence free survival of patients in our study was comparable to the reported literature but with significant lost to follow rate. Further large-scale, multi centre studies are needed to have a more comprehensive understanding of this heterogeneous disease in our population.

  8. Matrix-induced autologous chondrocyte implantation for the treatment of chondral defects of the knees in Chinese patients

    Directory of Open Access Journals (Sweden)

    Zhang ZW

    2014-12-01

    Full Text Available Zhongwen Zhang,1 Xin Zhong,2 Huiru Ji,1 Zibin Tang,1 Jianpeng Bai,1 Minmin Yao,1 Jianlei Hou,1 Minghao Zheng,3 David J Wood,3 Jiazhi Sun,4 Shu-Feng Zhou,4,5 Aibing Liu6 1Department of Orthopedics, General Hospital of Chinese People’s Armed Police Forces (CAPF, Beijing; 2Department of MRI Center, General Hospital of CAPF, Beijing, People’s Republic of China; 3Center for Orthopedic Research, School of Surgery and Pathology, University of Western Australia, Perth, Western Australia, Australia; 4Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA; 5Guizhou Provincial Key Laboratory for Regenerative Medicine, Stem Cell and Tissue Engineering Research Center and Sino–US Joint Laboratory for Medical Sciences, Guiyang Medical University, Guiyang, Guizhou; 6Medical Research Center, General Hospital of Chinese People’s Armed Police Forces (CAPF, Beijing, People’s Republic of China Abstract: Articular cartilage injury is the most common type of damage seen in clinical orthopedic practice. The matrix-induced autologous chondrocyte implant (MACI was developed to repair articular cartilage with an advance on the autologous chondrocyte implant procedure. This study aimed to evaluate whether MACI is a safe and efficacious cartilage repair treatment for patients with knee cartilage lesions. The primary outcomes were the Knee Injury and Osteoarthritis Outcome Score (KOOS domains and magnetic resonance imaging (MRI results, compared between baseline and postoperative months 3, 6, 12, and 24. A total of 15 patients (20 knees, with an average age of 33.9 years, had a mean defect size of 4.01 cm2. By 6-month follow-up, KOOS results demonstrated significant improvements in symptoms and knee-related quality of life. MRI showed significant improvements in four individual graft scoring parameters at 24 months postoperatively. At 24 months, 90% of MACI grafts had filled completely and 10% had good

  9. Osseous scintigraphy and auxiliary graft

    International Nuclear Information System (INIS)

    Khelifa, F.; Siles, S.; Puech, B.

    1992-01-01

    The scintigraphy could be a good way to survey the osseous graft: three cases are studied in which were recognized the presence of a graft, surinfection, graft lysis, pseudo-arthrosis, algodystrophy. 8 refs., 5 figs

  10. Meniscal allograft transplantation. Part 1: systematic review of graft biology, graft shrinkage, graft extrusion, graft sizing, and graft fixation.

    Science.gov (United States)

    Samitier, Gonzalo; Alentorn-Geli, Eduard; Taylor, Dean C; Rill, Brian; Lock, Terrence; Moutzouros, Vasilius; Kolowich, Patricia

    2015-01-01

    To provide a systematic review of the literature regarding five topics in meniscal allograft transplantation: graft biology, shrinkage, extrusion, sizing, and fixation. A systematic literature search was conducted using the PubMed (MEDLINE), ScienceDirect, and EBSCO-CINAHL databases. Articles were classified only in one topic, but information contained could be reported into other topics. Information was classified according to type of study (animal, in vitro human, and in vivo human) and level of evidence (for in vivo human studies). Sixty-two studies were finally included: 30 biology, 3 graft shrinkage, 11 graft extrusion, 17 graft size, and 6 graft fixation (some studies were categorized in more than one topic). These studies corresponded to 22 animal studies, 22 in vitro human studies, and 23 in vivo human studies (7 level II, 10 level III, and 6 level IV). The principal conclusions were as follows: (a) Donor cells decrease after MAT and grafts are repopulated with host cells form synovium; (b) graft preservation alters collagen network (deep freezing) and causes cell apoptosis with loss of viable cells (cryopreservation); (c) graft shrinkage occurs mainly in lyophilized and gamma-irradiated grafts (less with cryopreservation); (d) graft extrusion is common but has no clinical/functional implications; (e) overall, MRI is not superior to plain radiograph for graft sizing; (f) graft width size matching is more important than length size matching; (g) height appears to be the most important factor influencing meniscal size; (h) bone fixation better restores contact mechanics than suture fixation, but there are no differences for pullout strength or functional results; and (i) suture fixation has more risk of graft extrusion compared to bone fixation. Systematic review of level II-IV studies, Level IV.

  11. Advances in radiation grafting

    International Nuclear Information System (INIS)

    Hegazy, El-Sayed A.; AbdEl-Rehim, H.A.; Kamal, H.; Kandeel, K.A.

    2001-01-01

    Graft copolymerization is an attractive means for modifying base polymers because grafting frequently results in the superposition of properties relating to the backbone and pendent chains. Among the various methods for initiating the grafting reaction, ionizing radiation is the cleanest and most versatile method of grafting available. Ion-exchange membranes play an important role in modern technology, especially in separation and purification of materials. The search for improved membrane composition has considered almost every available polymeric material because of its great practical importance. Grafting of polymers with a mixture of monomers is important since different types of chains containing different functional groups are included. A great deal is focused on the waste treatment of heavy and toxic metals from wastewater because of the severe problems of environmental pollution. Functionalized polymers suitable for metal adsorption with their reactive functional groups such as carboxylic and pyridine groups suitable for waste treatment were prepared by radiation grafting method. More reactive chelating groups were further introduced to the grafted copolymer through its functional groups by chemical treatments with suitable reagents. The advances of radiation grafting and possible uses are briefly discussed

  12. Bone graft revascularization strategies

    NARCIS (Netherlands)

    Willems, W.F.

    2014-01-01

    Reconstruction of avascular necrotic bone by pedicled bone grafting is a well-known treatment with little basic research supporting its application. A new canine model was used to simulate carpal bone avascular necrosis. Pedicled bone grafting proved to increase bone remodeling and bone blood flow,

  13. Reduction of Environmental Temperature Mitigates Local Anesthetic Cytotoxicity in Bovine Articular Chondrocytes

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    Tarik Onur, Alexis Dang

    2014-09-01

    Full Text Available The purpose of this study was to assess whether reducing environmental temperature will lead to increased chondrocyte viability following injury from a single-dose of local anesthetic treatment. Bovine articular chondrocytes from weight bearing portions of femoral condyles were harvested and cultured. 96-well plates were seeded with 15,000 chondrocytes per well. Chondrocytes were treated with one of the following conditions: ITS Media, 1x PBS, 2% lidocaine, 0.5% bupivacaine, or 0.5% ropivacaine. Each plate was then incubated at 37°C, 23°C, or 4°C for one hour and then returned to media at 37°C. Chondrocyte viability was assessed 24 hours after treatment. Chondrocyte viability is presented as a ratio of the fluorescence of the treatment group over the average of the media group at that temperature (ratio ± SEM. At 37°C, lidocaine (0.35 ± 0.04 and bupivacaine (0.30 ± 0.05 treated chondrocytes show low cell viability when compared to the media (1.00 ± 0.03 control group (p < 0.001. Lidocaine treated chondrocytes were significantly more viable at 23°C (0.84 ± 0.08 and 4°C (0.86±0.085 than at 37°C (p < 0.001. Bupivacaine treated chondrocytes were significantly more viable at 4°C (0.660 ± 0.073 than at 37°C or 23°C (0.330 ± 0.069 (p < 0.001 and p = 0.002 respectively. Reducing the temperature from 37°C to 23°C during treatment with lidocaine increases chondrocyte viability following injury. Chondrocytes treated with bupivacaine can be rescued by reducing the temperature to 4°C.

  14. SHP2 regulates chondrocyte terminal differentiation, growth plate architecture and skeletal cell fates.

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    Margot E Bowen

    Full Text Available Loss of PTPN11/SHP2 in mice or in human metachondromatosis (MC patients causes benign cartilage tumors on the bone surface (exostoses and within bones (enchondromas. To elucidate the mechanisms underlying cartilage tumor formation, we investigated the role of SHP2 in the specification, maturation and organization of chondrocytes. Firstly, we studied chondrocyte maturation by performing RNA-seq on primary chondrocyte pellet cultures. We found that SHP2 depletion, or inhibition of the ERK1/2 pathway, delays the terminal differentiation of chondrocytes from the early-hypertrophic to the late-hypertrophic stage. Secondly, we studied chondrocyte maturation and organization in mice with a mosaic postnatal inactivation of Ptpn11 in chondrocytes. We found that the vertebral growth plates of these mice have expanded domains of early-hypertrophic chondrocytes that have not yet terminally differentiated, and their enchondroma-like lesions arise from chondrocytes displaced from the growth plate due to a disruption in the organization of maturation and ossification zones. Furthermore, we observed that lesions from human MC patients also display disorganized chondrocyte maturation zones. Next, we found that inactivation of Ptpn11 in Fsp1-Cre-expressing fibroblasts induces exostosis-like outgrowths, suggesting that loss of SHP2 in cells on the bone surface and at bone-ligament attachment sites induces ectopic chondrogenesis. Finally, we performed lineage tracing to show that exostoses and enchondromas in mice likely contain mixtures of wild-type and SHP2-deficient chondrocytes. Together, these data indicate that in patients with MC, who are heterozygous for inherited PTPN11 loss-of-function mutations, second-hit mutations in PTPN11 can induce enchondromas by disrupting the organization and delaying the terminal differentiation of growth plate chondrocytes, and can induce exostoses by causing ectopic chondrogenesis of cells on the bone surface. Furthermore, the

  15. [Preference Changes Regarding Future Work Area and Intended Position Among German Residents after Four Years of Residency].

    Science.gov (United States)

    Ziegler, Stine; van den Bussche, Hendrik; Römer, Farina; Krause-Solberg, Lea; Scherer, Martin

    2017-06-01

    Introduction  We investigated the preferences of medical residents in Germany with regard to future working place (hospital or private practice) and position (employment/self-employment in private practice; resp. specialist/senior or chief physician in the hospital). This is analysed in a gender comparative perspective, including the influence of parenthood. Methods  Annual postal surveys among graduates of seven medical faculties in Germany from their last year ("Practical Year") until after four years of postgraduate training. The return rate at baseline was 48 % and the four surveys after reached rates from 85 % up. In all samples about two thirds were women, which corresponds to the actual gender differentiation in under- and postgraduate training. Descriptive statistics and regression analyses were performed. Results  Compared to private practice the hospital is clearly preferred, although the attraction of hospital jobs decreased over the years. The decision for or against the hospital is connected to the discipline. Working in private practice is seen as possibility for part time work. Men prefer self-employment whereas women prefer to work under an employment contract. In the hospital, male doctors prefer to work in leading positions. Those positions are associated with full-time work. Leadership training especially takes place in university hospitals. Discussion  Three trends are recognized: Reluctance against leading positions, growing interest for part time work and rising popularity of work as an employee in private practice. Those trends can be understood as a rejection of traditional professional role models. The realization of these preferences is easily feasible because of the current labour market situation. Therefore, emerging problems have to be faced in another way. A change of gender-typical role models was rarely detected. © Georg Thieme Verlag KG Stuttgart · New York.

  16. Experiences of internet-delivered cognitive behavior therapy for social anxiety disorder four years later: A qualitative study

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    Camilla Olsson Halmetoja

    2014-07-01

    Full Text Available The current study is a qualitative follow-up of a study on guided internet-delivered cognitive behavior therapy (ICBT for social anxiety disorder (SAD, conducted four years after treatment completion. The main aim was to capture participants' description of their experiences of the treatment, their view on treatment effects, memories of the treatment, and whether they continued using the gained knowledge after treatment. Sixty participants were selected from the original study's treatment group. A criterion based sampling approach was used based on the obtained treatment effect, and with a minimum of five completed treatment modules. E-mail invitations were sent, with information about the follow-up and the instruction to respond if interested in participating. Twelve semi-structured interviews were made and the material was analyzed using an approach based on grounded theory. The results showed that all participants found the treatment to have some effect, but they also found it to be demanding, difficult, and hard. Many appreciated to hear of the experiences of other participants in the online forum. Under the theme of memory, most could describe the setup of the treatment in general terms. The exposure module was mentioned by all, cognitive restructuring by most, and some also reported memories of the psychoeducation. A core process was identified which involved how the attained treatment effect was viewed over the time, and how this view changed from treatment completion to current time. The findings outlined in this study describe how treatment effects can be sustained via an active approach to the treatment and the symptoms of SAD.

  17. Shift work schedule and night work load: Effects on body mass index - a four-year longitudinal study.

    Science.gov (United States)

    Buchvold, Hogne Vikanes; Pallesen, Ståle; Waage, Siri; Bjorvatn, Bjørn

    2018-05-01

    Objectives The aim of this study was to investigate changes in body mass index (BMI) between different work schedules and different average number of yearly night shifts over a four-year follow-up period. Methods A prospective study of Norwegian nurses (N=2965) with different work schedules was conducted: day only, two-shift rotation (day and evening shifts), three-shift rotation (day, evening and night shifts), night only, those who changed towards night shifts, and those who changed away from schedules containing night shifts. Paired student's t-tests were used to evaluate within subgroup changes in BMI. Multiple linear regression analysis was used to evaluate between groups effects on BMI when adjusting for BMI at baseline, sex, age, marital status, children living at home, and years since graduation. The same regression model was used to evaluate the effect of average number of yearly night shifts on BMI change. Results We found that night workers [mean difference (MD) 1.30 (95% CI 0.70-1.90)], two shift workers [MD 0.48 (95% CI 0.20-0.75)], three shift workers [MD 0.46 (95% CI 0.30-0.62)], and those who changed work schedule away from [MD 0.57 (95% CI 0.17-0.84)] or towards night work [MD 0.63 (95% CI 0.20-1.05)] all had significant BMI gain (Pnight workers had significantly larger BMI gain compared to day workers [B=0.89 (95% CI 0.06-1.72), Pnight shifts and BMI change using our multiple linear regression model. Conclusions After adjusting for possible confounders, we found that BMI increased significantly more among night workers compared to day workers.

  18. Four-year follow-up study in a NF1 boy with a focal pontine hamartoma.

    Science.gov (United States)

    Parisi, Pasquale; Persechino, Severino; Paolino, Maria Chiara; Nicita, Francesco; Torrente, Isabella; Bozzao, Alessandro; Villa, Maria Pia

    2013-02-11

    Neurofibromatosis is a collective name for a group of genetic conditions in which benign tumours affect the nervous system. Type 1 is caused by a genetic mutation in the NF1 gene (OMIM 613113) and symptoms can vary dramatically between individuals, even within the same family. Some people have very mild skin changes, whereas others suffer severe medical complications. The condition usually appears in childhood and is diagnosed if two of the following are present: six or more café-au-lait patches larger than 1.5 cm in diameter, axillary or groin freckling, 2 or more Lisch nodules (small pigmented areas in the iris of the eye), 2 or more neurofibromas, optic pathway gliomas, bone dysplasia, and a first-degree family relative with Neurofibromatosis type 1. The pattern of inheritance is autosomal dominant, however, half of all NF1 cases are 'sporadic' and there is no family history. Neurofibromatosis type 1 is an extremely variable condition whose morbidity and mortality is largely dictated by the occurrence of the many complications that may involve any of the body systems. We describe a family affected by NF1 in whom genetic molecular analysis identified the same mutation in the son and father. Routine MRI showed pontine focal lesions in the eight-year-old son, though not in the father. We performed a four years follow-up study and at follow-up pontine hamartoma size remained unchanged in the son, and the father showed still no brain lesions, confirming thus an intra-familial phenotype variability.

  19. Four-year follow-up study in a NF1 Boy with a focal pontine hamartoma

    Directory of Open Access Journals (Sweden)

    Parisi Pasquale

    2013-02-01

    Full Text Available Abstract Neurofibromatosis is a collective name for a group of genetic conditions in which benign tumours affect the nervous system. Type 1 is caused by a genetic mutation in the NF1 gene (OMIM 613113 and symptoms can vary dramatically between individuals, even within the same family. Some people have very mild skin changes, whereas others suffer severe medical complications. The condition usually appears in childhood and is diagnosed if two of the following are present: six or more café-au-lait patches larger than 1.5 cm in diameter, axillary or groin freckling, 2 or more Lisch nodules (small pigmented areas in the iris of the eye, 2 or more neurofibromas, optic pathway gliomas, bone dysplasia, and a first-degree family relative with Neurofibromatosis type 1. The pattern of inheritance is autosomal dominant, however, half of all NF1 cases are ‘sporadic’ and there is no family history. Neurofibromatosis type 1 is an extremely variable condition whose morbidity and mortality is largely dictated by the occurrence of the many complications that may involve any of the body systems. We describe a family affected by NF1 in whom genetic molecular analysis identified the same mutation in the son and father. Routine MRI showed pontine focal lesions in the eight-year-old son, though not in the father. We performed a four years follow-up study and at follow-up pontine hamartoma size remained unchanged in the son, and the father showed still no brain lesions, confirming thus an intra-familial phenotype variability.

  20. Clinical and Hematological Evaluation of Patients with Sickle Cell Anemia Before and After Four Years of Using Hydroxyurea

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    Ieda Maria Gonçalves Pacce Bispo

    2017-06-01

    Full Text Available Objective: Evaluating clinical and hematological-clinical parameters of patients with sickle cell anemia (SCA before and after four years of using hydroxyurea (HU.  Method: A retrospective cohort study implementing a quantitative, descriptive and analytical approach developed in two public teaching hospitals located in the Central-West region of Brazil, from November 2010 to October 2011. Data collection was performed through medical records of 32 patients with SCA to assess clinical and hematological parameters before and after HU treatment. The study was approved by the UFMS Ethics Committee under protocol number 1890/2010. Results: All of the 32 patients were homozygous with a mean age in the prescription of hydroxyurea of 19.72±7.58 years, an initial dose of 15.59±4.27 mg/kg/day, and 22.48±5.35 mg/kg/day in the fourth year of treatment. Regarding the use of HU, average values of some hematological parameters presented a significant difference in the fourth year compared to the mean values prior to HU use, such as fetal hemoglobin (14.49±7.52%, red blood cells (2.54±0.38x1012/L, hematocrit (25.30±4.03% and hemoglobin (9.22±3.34g/dL.  Conclusion: Treatment with hydroxyurea showed a significant increase in fetal hemoglobin levels, increased hemoglobin, hematocrit and average corpuscular hemoglobin concentration, with reduced episodes of pain, infection and acute chest syndrome in such a way as to reaffirm its efficiency in treating these patients. Keywords: Hemoglobin; Sickle Cell Anemia; Hydroxyurea.

  1. Drought, Frost, Rain and Sunshine. Four Years of Sap Flow Measurements for One of the World's Largest Conifers

    Science.gov (United States)

    Macinnis-Ng, C.; Taylor, D. T.; Kaplick, J.; Clearwater, M.

    2015-12-01

    Amongst the largest and longest lived conifers in the world, the endemic New Zealand kauri, Agathis australis, provides a proxy-climate record dating back 4000 y. Tree-ring widths provide a strong indicator of the occurrence of El Niño Southern Oscillation (ENSO) events. We are measuring physiological processes, including carbon uptake and loss, leaf-scale gas exchange and sap flow together with meteorological data to explore the mechanisms of the climate response of this iconic and culturally significant species. In this continuous 15 min time interval sap flow dataset spanning four years, we have captured very wet and very dry summer periods. Winter flow rates peaked lower than summer flow rates and winter flow also started later and finished earlier in the day, resulting in less water use. Larger, canopy dominant trees (DBH up to 176 cm) had large sapwood area (sapwood depth up to 18 cm) and faster flow rates and therefore dominated stand water use. During dry periods, smaller trees (DBH 20-80 cm) were more responsive to dry soils than larger trees, suggesting access to deeper soil water stores. Leaf-scale gas exchange rates were low with very low stomatal conductance values reflecting known vulnerability to xylem embolism. Night-time refilling of sapwood was particularly evident during the summer drought with evidence that refilling was incomplete as the drought progressed. Photosynthetically active radiation and vapour pressure deficit are strongly correlated with sap flow across all seasons, a promising indicator for future modelling work on this dataset. Water saving strategies and stand-scale water budgets are discussed.

  2. Carriage of Staphylococcus aureus among Portuguese nursing students: A longitudinal cohort study over four years of education.

    Directory of Open Access Journals (Sweden)

    Teresa Conceição

    Full Text Available Staphylococcus aureus is a major human pathogen that can colonize healthy people mainly in the anterior nares. The aim of the present study was to evaluate S. aureus nasal colonization over time among Portuguese nursing students, including methicillin-resistant S. aureus (MRSA.In this longitudinal cohort study, we collected 280 nasal swabs from nursing students at 14 time points over four years of schooling (2012-2016. The isolates were characterized by pulsed-field gel electrophoresis (PFGE, spa typing, multilocus sequence typing (MLST, and SCCmec typing for MRSA. Among 47 students, 20 (43% carried methicillin-susceptible S. aureus (MSSA at admission, but none was colonized with MRSA. A total of 19 students (40% became colonized after exposure during the nursing training, out of which five carried MRSA. Overall, 39 students (83% had S. aureus detected at least once during the study period. Among the 97 MSSA isolates, most (65% belonged to four clones: PFGE A-ST30 (21%, B-ST72 (20%, C-ST508 (13%, and D-ST398 (11%. Three of the five MRSA carriers were colonized with the predominant clone circulating in Portuguese hospitals (ST22-IVh and two with ST3162-II. Colonization of nursing students was highly dynamic with continuous appearance of strains with distinct PFGE types in the same individual.A considerable proportion of students became colonized by S. aureus, including MRSA, during the nursing education, evidencing this population represents an important reservoir of S. aureus. Therefore, education on infection control measures in nursing schools is of major importance.

  3. Atf4 regulates chondrocyte proliferation and differentiation during endochondral ossification by activating Ihh transcription.

    Science.gov (United States)

    Wang, Weiguang; Lian, Na; Li, Lingzhen; Moss, Heather E; Wang, Weixi; Perrien, Daniel S; Elefteriou, Florent; Yang, Xiangli

    2009-12-01

    Activating transcription factor 4 (Atf4) is a leucine-zipper-containing protein of the cAMP response element-binding protein (CREB) family. Ablation of Atf4 (Atf4(-/-)) in mice leads to severe skeletal defects, including delayed ossification and low bone mass, short stature and short limbs. Atf4 is expressed in proliferative and prehypertrophic growth plate chondrocytes, suggesting an autonomous function of Atf4 in chondrocytes during endochondral ossification. In Atf4(-/-) growth plate, the typical columnar structure of proliferative chondrocytes is disturbed. The proliferative zone is shortened, whereas the hypertrophic zone is transiently expanded. The expression of Indian hedgehog (Ihh) is markedly decreased, whereas the expression of other chondrocyte marker genes, such as type II collagen (Col2a1), PTH/PTHrP receptor (Pth1r) and type X collagen (Col10a1), is normal. Furthermore, forced expression of Atf4 in chondrocytes induces endogenous Ihh mRNA, and Atf4 directly binds to the Ihh promoter and activates its transcription. Supporting these findings, reactivation of Hh signaling pharmacologically in mouse limb explants corrects the Atf4(-/-) chondrocyte proliferation and short limb phenotypes. This study thus identifies Atf4 as a novel transcriptional activator of Ihh in chondrocytes that paces longitudinal bone growth by controlling growth plate chondrocyte proliferation and differentiation.

  4. Curcumin Inhibits Chondrocyte Hypertrophy of Mesenchymal Stem Cells through IHH and Notch Signaling Pathways.

    Science.gov (United States)

    Cao, Zhen; Dou, Ce; Dong, Shiwu

    2017-01-01

    Using tissue engineering technique to repair cartilage damage caused by osteoarthritis is a promising strategy. However, the regenerated tissue usually is fibrous cartilage, which has poor mechanical characteristics compared to hyaline cartilage. Chondrocyte hypertrophy plays an important role in this process. Thus, it is very important to find out a suitable way to maintain the phenotype of chondrocytes and inhibit chondrocyte hypertrophy. Curcumin deriving from turmeric was reported with anti-inflammatory and anti-tumor pharmacological effects. However, the role of curcumin in metabolism of chondrocytes, especially in the chondrocyte hypertrophy remains unclear. Mesenchymal stem cells (MSCs) are widely used in cartilage tissue engineering as seed cells. So we investigated the effect of curcumin on chondrogenesis and chondrocyte hypertrophy in MSCs through examination of cell viability, glycosaminoglycan synthesis and specific gene expression. We found curcumin had no effect on expression of chondrogenic markers including Sox9 and Col2a1 while hypertrophic markers including Runx2 and Col10a1 were down-regulated. Further exploration showed that curcumin inhibited chondrocyte hypertrophy through Indian hedgehog homolog (IHH) and Notch signalings. Our results indicated curcumin was a potential agent in modulating cartilage homeostasis and maintaining chondrocyte phenotype.

  5. Nicotine promotes proliferation and collagen synthesis of chondrocytes isolated from normal human and osteoarthritis patients.

    Science.gov (United States)

    Ying, Xiaozhou; Cheng, Shaowen; Shen, Yue; Cheng, Xiaojie; An Rompis, Ferdinand; Wang, Wei; Lin, Zhongqin; Chen, Qingyu; Zhang, Wei; Kou, Dongquan; Peng, Lei; Tian, Xin Qiao; Lu, Chuan Zhu

    2012-01-01

    The aims of the study were to show the direct effect of nicotine with different concentrations (0, 25, 50, and 100 ng/ml) on chondrocytes isolated from normal human and osteoarthritis patients, respectively. Microscopic observation was performed during the culture with an inverted microscope. Methyl thiazolyl tetrazolium (MTT) assay method was adopted to observe the influence of nicotine on the proliferation of chondrocytes, and real-time PCR and ELISA were used to assay the mRNA and protein expression of type II collagen and aggrecan, respectively. We discovered that the OA chondrocytes were similar to fibroblasts in shape and grow slower than normal chondrocytes. The proliferation of the two kinds of chondrocytes was increased in a concentration-dependent manner and in a time-dependent manner (P<0.05). Also, we found that the mRNA level of type II collagen were upregulated under 25-100 ng/ml nicotine doses both in the two kinds of chondrocytes compared with control. The expression of protein levels of type II collagen were synthesized in line with the increase in mRNA. No effect was observed on aggrecan synthesis with any nicotine dose. We concluded that nicotine has the same effect on both chondrocytes, obtained either from osteoarthritis patients or from normal human, and the positive effect of smoking in OA may relate to the alteration in metabolism of chondrocytes.

  6. Modulation of Hyaluronan Synthesis by the Interaction between Mesenchymal Stem Cells and Osteoarthritic Chondrocytes

    Directory of Open Access Journals (Sweden)

    Eliane Antonioli

    2015-01-01

    Full Text Available Bone marrow mesenchymal stem cells (BM-MSCs are considered a good source for cellular therapy in cartilage repair. But, their potential to repair the extracellular matrix, in an osteoarthritic environment, is still controversial. In osteoarthritis (OA, anti-inflammatory action and extracellular matrix production are important steps for cartilage healing. This study examined the interaction of BM-MSC and OA-chondrocyte on the production of hyaluronan and inflammatory cytokines in a Transwell system. We compared cocultured BM-MSCs and OA-chondrocytes with the individually cultured controls (monocultures. There was a decrease in BM-MSCs cell count in coculture with OA-chondrocytes when compared to BM-MSCs alone. In monoculture, BM-MSCs produced higher amounts of hyaluronan than OA-chondrocytes and coculture of BM-MSCs with OA-chondrocytes increased hyaluronan production per cell. Hyaluronan synthase-1 mRNA expression was upregulated in BM-MSCs after coculture with OA-chondrocytes, whereas hyaluronidase-1 was downregulated. After coculture, lower IL-6 levels were detected in BM-MSCs compared with OA-chondrocytes. These results indicate that, in response to coculture with OA-chondrocytes, BM-MSCs change their behavior by increasing production of hyaluronan and decreasing inflammatory cytokines. Our results indicate that BM-MSCs per se could be a potential tool for OA regenerative therapy, exerting short-term effects on the local microenvironment even when cell:cell contact is not occurring.

  7. Engineering Cartilage Tissue by Pellet Coculture of Chondrocytes and Mesenchymal Stromal Cells

    NARCIS (Netherlands)

    Wu, Ling; Post, Janine Nicole; Karperien, Hermanus Bernardus Johannes; Westendorf, Jennifer J.; van Wijnen, Andre J.

    2015-01-01

    Coculture of chondrocytes and mesenchymal stromal cells (MSCs) in pellets has been shown to be beneficial in engineering cartilage tissue in vitro. In these cultures trophic effects of MSCs increase the proliferation and matrix deposition of chondrocytes. Thus, large cartilage constructs can be made

  8. RAGE and activation of chondrocytes and fibroblast-like synoviocytes in joint diseases

    NARCIS (Netherlands)

    Steenvoorden, Marjan Maria Claziena

    2007-01-01

    This dissertation describes a new model in which cartilage degradation can be studied. New cartilage is formed by bovine chondrocytes obtained from the slaughterhouse and cocultured with synovial cells from rheumatoid arthritis (RA) patients to study the interaction between the chondrocytes and

  9. High throughput generated micro-aggregates of chondrocytes stimulate cartilage formation in vitro and in vivo

    NARCIS (Netherlands)

    Moreira Teixeira, Liliana; Leijten, Jeroen Christianus Hermanus; Sobral, J.; Jin, R.; van Apeldoorn, Aart A.; Feijen, Jan; van Blitterswijk, Clemens; Dijkstra, Pieter J.; Karperien, Hermanus Bernardus Johannes

    2012-01-01

    Cell-based cartilage repair strategies such as matrix-induced autologous chondrocyte implantation (MACI) could be improved by enhancing cell performance. We hypothesised that micro-aggregates of chondrocytes generated in high-throughput prior to implantation in a defect could stimulate cartilaginous

  10. Spatiotemporal Determinants of Urban Leptospirosis Transmission: Four-Year Prospective Cohort Study of Slum Residents in Brazil.

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    José E Hagan

    2016-01-01

    Full Text Available Rat-borne leptospirosis is an emerging zoonotic disease in urban slum settlements for which there are no adequate control measures. The challenge in elucidating risk factors and informing approaches for prevention is the complex and heterogeneous environment within slums, which vary at fine spatial scales and influence transmission of the bacterial agent.We performed a prospective study of 2,003 slum residents in the city of Salvador, Brazil during a four-year period (2003-2007 and used a spatiotemporal modelling approach to delineate the dynamics of leptospiral transmission. Household interviews and Geographical Information System surveys were performed annually to evaluate risk exposures and environmental transmission sources. We completed annual serosurveys to ascertain leptospiral infection based on serological evidence. Among the 1,730 (86% individuals who completed at least one year of follow-up, the infection rate was 35.4 (95% CI, 30.7-40.6 per 1,000 annual follow-up events. Male gender, illiteracy, and age were independently associated with infection risk. Environmental risk factors included rat infestation (OR 1.46, 95% CI, 1.00-2.16, contact with mud (OR 1.57, 95% CI 1.17-2.17 and lower household elevation (OR 0.92 per 10m increase in elevation, 95% CI 0.82-1.04. The spatial distribution of infection risk was highly heterogeneous and varied across small scales. Fixed effects in the spatiotemporal model accounted for the majority of the spatial variation in risk, but there was a significant residual component that was best explained by the spatial random effect. Although infection risk varied between years, the spatial distribution of risk associated with fixed and random effects did not vary temporally. Specific "hot-spots" consistently had higher transmission risk during study years.The risk for leptospiral infection in urban slums is determined in large part by structural features, both social and environmental. Our findings

  11. PKS 2005-489 at VHE: four years of monitoring with HESS and simultaneous multi-wavelength observations

    Science.gov (United States)

    H.E.S.S. Collaboration; Acero, F.; Aharonian, F.; Akhperjanian, A. G.; Anton, G.; Barres de Almeida, U.; Bazer-Bachi, A. R.; Becherini, Y.; Behera, B.; Benbow, W.; Bernlöhr, K.; Bochow, A.; Boisson, C.; Bolmont, J.; Borrel, V.; Brucker, J.; Brun, F.; Brun, P.; Bühler, R.; Bulik, T.; Büsching, I.; Boutelier, T.; Chadwick, P. M.; Charbonnier, A.; Chaves, R. C. G.; Cheesebrough, A.; Chounet, L.-M.; Clapson, A. C.; Coignet, G.; Costamante, L.; Dalton, M.; Daniel, M. K.; Davids, I. D.; Degrange, B.; Deil, C.; Dickinson, H. J.; Djannati-Ataï, A.; Domainko, W.; O'C. Drury, L.; Dubois, F.; Dubus, G.; Dyks, J.; Dyrda, M.; Egberts, K.; Eger, P.; Espigat, P.; Fallon, L.; Farnier, C.; Fegan, S.; Feinstein, F.; Fiasson, A.; Förster, A.; Fontaine, G.; Füßling, M.; Gabici, S.; Gallant, Y. A.; Gérard, L.; Gerbig, D.; Giebels, B.; Glicenstein, J. F.; Glück, B.; Goret, P.; Göring, D.; Hauser, M.; Heinz, S.; Heinzelmann, G.; Henri, G.; Hermann, G.; Hinton, J. A.; Hoffmann, A.; Hofmann, W.; Hofverberg, P.; Holleran, M.; Hoppe, S.; Horns, D.; Jacholkowska, A.; de Jager, O. C.; Jahn, C.; Jung, I.; Katarzyński, K.; Katz, U.; Kaufmann, S.; Kerschhaggl, M.; Khangulyan, D.; Khélifi, B.; Keogh, D.; Klochkov, D.; Kluźniak, W.; Kneiske, T.; Komin, Nu.; Kosack, K.; Kossakowski, R.; Lamanna, G.; Lenain, J.-P.; Lohse, T.; Marandon, V.; Martineau-Huynh, O.; Marcowith, A.; Masbou, J.; Maurin, D.; McComb, T. J. L.; Medina, M. C.; Méhault, J.; Moderski, R.; Moulin, E.; Naumann-Godo, M.; de Naurois, M.; Nedbal, D.; Nekrassov, D.; Nicholas, B.; Niemiec, J.; Nolan, S. J.; Ohm, S.; Olive, J.-F.; de Oña Wilhelmi, E.; Orford, K. J.; Ostrowski, M.; Panter, M.; Paz Arribas, M.; Pedaletti, G.; Pelletier, G.; Petrucci, P.-O.; Pita, S.; Pühlhofer, G.; Punch, M.; Quirrenbach, A.; Raubenheimer, B. C.; Raue, M.; Rayner, S. M.; Renaud, M.; Rieger, F.; Ripken, J.; Rob, L.; Rosier-Lees, S.; Rowell, G.; Rudak, B.; Rulten, C. B.; Ruppel, J.; Sahakian, V.; Santangelo, A.; Schlickeiser, R.; Schöck, F. M.; Schwanke, U.; Schwarzburg, S.; Schwemmer, S.; Shalchi, A.; Sikora, M.; Skilton, J. L.; Sol, H.; Stawarz, Ł.; Steenkamp, R.; Stegmann, C.; Stinzing, F.; Superina, G.; Szostek, A.; Tam, P. H.; Tavernet, J.-P.; Terrier, R.; Tibolla, O.; Tluczykont, M.; van Eldik, C.; Vasileiadis, G.; Venter, C.; Venter, L.; Vialle, J. P.; Vincent, P.; Vivier, M.; Völk, H. J.; Volpe, F.; Wagner, S. J.; Ward, M.; Zdziarski, A. A.; Zech, A.

    2010-02-01

    Aims: Our aim is to study the very high energy (VHE; E>100 GeV) γ-ray emission from BL Lac objects and the evolution in time of their broad-band spectral energy distribution (SED). Methods: VHE observations of the high-frequency peaked BL Lac object PKS 2005-489 were made with the High Energy Stereoscopic System (HESS) from 2004 through 2007. Three simultaneous multi-wavelength campaigns at lower energies were performed during the HESS data taking, consisting of several individual pointings with the XMM-Newton and RXTE satellites. Results: A strong VHE signal, ~17σ total, from PKS 2005-489 was detected during the four years of HESS observations (90.3 h live time). The integral flux above the average analysis threshold of 400 GeV is ~3% of the flux observed from the Crab Nebula and varies weakly on time scales from days to years. The average VHE spectrum measured from ~300 GeV to ~5 TeV is characterized by a power law with a photon index, Γ = 3.20± 0.16_stat± 0.10_syst. At X-ray energies the flux is observed to vary by more than an order of magnitude between 2004 and 2005. Strong changes in the X-ray spectrum (ΔΓX ≈ 0.7) are also observed, which appear to be mirrored in the VHE band. Conclusions: The SED of PKS 2005-489, constructed for the first time with contemporaneous data on both humps, shows significant evolution. The large flux variations in the X-ray band, coupled with weak or no variations in the VHE band and a similar spectral behavior, suggest the emergence of a new, separate, harder emission component in September 2005. Supported by CAPES Foundation, Ministry of Education of Brazil.Now at Harvard-Smithsonian Center for Astrophysics, Cambridge, USA.Now at W.W. Hansen Experimental Physics Laboratory & Kavli Institute for Particle Astrophysics and Cosmology, Stanford University, Stanford, USA.

  12. Spatiotemporal Determinants of Urban Leptospirosis Transmission: Four-Year Prospective Cohort Study of Slum Residents in Brazil.

    Science.gov (United States)

    Hagan, José E; Moraga, Paula; Costa, Federico; Capian, Nicolas; Ribeiro, Guilherme S; Wunder, Elsio A; Felzemburgh, Ridalva D M; Reis, Renato B; Nery, Nivison; Santana, Francisco S; Fraga, Deborah; Dos Santos, Balbino L; Santos, Andréia C; Queiroz, Adriano; Tassinari, Wagner; Carvalho, Marilia S; Reis, Mitermayer G; Diggle, Peter J; Ko, Albert I

    2016-01-01

    Rat-borne leptospirosis is an emerging zoonotic disease in urban slum settlements for which there are no adequate control measures. The challenge in elucidating risk factors and informing approaches for prevention is the complex and heterogeneous environment within slums, which vary at fine spatial scales and influence transmission of the bacterial agent. We performed a prospective study of 2,003 slum residents in the city of Salvador, Brazil during a four-year period (2003-2007) and used a spatiotemporal modelling approach to delineate the dynamics of leptospiral transmission. Household interviews and Geographical Information System surveys were performed annually to evaluate risk exposures and environmental transmission sources. We completed annual serosurveys to ascertain leptospiral infection based on serological evidence. Among the 1,730 (86%) individuals who completed at least one year of follow-up, the infection rate was 35.4 (95% CI, 30.7-40.6) per 1,000 annual follow-up events. Male gender, illiteracy, and age were independently associated with infection risk. Environmental risk factors included rat infestation (OR 1.46, 95% CI, 1.00-2.16), contact with mud (OR 1.57, 95% CI 1.17-2.17) and lower household elevation (OR 0.92 per 10m increase in elevation, 95% CI 0.82-1.04). The spatial distribution of infection risk was highly heterogeneous and varied across small scales. Fixed effects in the spatiotemporal model accounted for the majority of the spatial variation in risk, but there was a significant residual component that was best explained by the spatial random effect. Although infection risk varied between years, the spatial distribution of risk associated with fixed and random effects did not vary temporally. Specific "hot-spots" consistently had higher transmission risk during study years. The risk for leptospiral infection in urban slums is determined in large part by structural features, both social and environmental. Our findings indicate that

  13. GeoFORCE Alaska: Four-Year Field Program Brings Rural Alaskan High School Students into the STEM Pipeline

    Science.gov (United States)

    Fowell, S. J.; Rittgers, A.; Stephens, L.; Hutchinson, S.; Peters, H.; Snow, E.; Wartes, D.

    2016-12-01

    GeoFORCE Alaska is a four-year, field-based, summer geoscience program designed to raise graduation rates in rural Alaskan high schools, encourage participants to pursue college degrees, and increase the diversity of Alaska's technical workforce. Residents of predominantly Alaska Native villages holding degrees in science, technology, engineering, or math (STEM) bring valuable perspectives to decisions regarding management of cultural and natural resources. However, between 2010 and 2015 the average dropout rate for students in grades 7-12 was 8.5% per year in the North Slope School District and 7% per year in the Northwest Arctic School District. 2015 graduation rates were 70% and 75%, respectively. Statewide statistics highlight the challenge for Alaska Native students. During the 2014-2015 school year alone 37.6% of Alaska Native students dropped out of Alaskan public schools. At the college level, Alaska Native students are underrepresented in University of Alaska Fairbanks (UAF) science departments. Launched in 2012 by UAF in partnership with the longstanding University of Texas at Austin program, GeoFORCE applies the cohort model, leading the same group of high school students on geological field academies during four consecutive summers. Through a combination of active learning, teamwork, and hands-on projects at spectacular geological locations, students gain academic skills and confidence that facilitate high school and college success. To date, GeoFORCE Alaska has recruited two cohorts. 78% of these students identify as Alaska Native, reflecting community demographics. The inaugural cohort of 18 students from the North Slope Borough completed the Fourth-Year Academy in summer 2015. 94% of these students graduated from high school, at least 72% plan to attend college, and 33% will major in geoscience. A second cohort of 34 rising 9th and 10th graders entered the program in 2016. At the request of corporate sponsors, this cohort was recruited from both the

  14. Overview of the SEU project for extended burnup at the Atucha-I NPP. Four years of operating experience

    International Nuclear Information System (INIS)

    Fink, J.M.; Higa, M.; Perez, R.; Pineyro, J.; Sidelnik, J.; Casario, J.A.; Alvarez, L.

    2002-01-01

    Atucha I is a 357 MWe nuclear station moderated and cooled with heavy water, of German design located in Argentina. Fuelling is on-power and the plant was originally fuelled with natural uranium. To reduce fuel costs a program was initiated in August 1993 to introduce gradually slightly enriched uranium (SEU) fuel (0.85 w% U-235) with an associated burnup increase from 5900 MWd/tU to 11300 MWd/tU. The introduction of SEU fuel started in January 1995 and the program was divided in three Phases with an upper limit of SEU FA in the core: 12, 60 and 252 (full core) and licensing documentation was prepared for each Phase. This paper describes the most important aspects of the operating and project experience, and some factors limiting the burnup extension from an operation point of view. After four years of the program and with 181 SEU FA (71%) of the core, the operating experience has been good and without unfavourable effects due to the use of SEU fuel with the only exception of a small increase of the time to reach full power in plant startups or power cycling. In particular, the new criteria to prevent PCI failures in power ramps for higher burnup SEU fuel in refueling operations, plant startups or power cycling has been effective. The average discharge burnup of the SEU fuel taken out of the reactor in 1998 was 11263 MWd/tU. The average discharge burnup of the natural fuel in the same year was 6640 MWd/tU, with an increase of about 12% of the original value for a natural fuel core. The average number of fresh fuel assemblies per full power day was being reduced from 1.31 to 0.92 in 1998 and 0.83 in 1999. The fuel costs dropped gradually during the program from 9.38 (with natural uranium fuel) to 6.57 $/MWh in the first four months of 1999 (taking as reference the NU and SEU FA costs for 1999). Because of this the SEU program has been an important contribution to the reduction of Atucha I operating costs and to the competitiveness of nuclear power generation against

  15. 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.

  16. R-spondin 2 facilitates differentiation of proliferating chondrocytes into hypertrophic chondrocytes by enhancing Wnt/β-catenin signaling in endochondral ossification

    International Nuclear Information System (INIS)

    Takegami, Yasuhiko; Ohkawara, Bisei; Ito, Mikako; Masuda, Akio; Nakashima, Hiroaki; Ishiguro, Naoki; Ohno, Kinji

    2016-01-01

    Endochondral ossification is a crucial process for longitudinal growth of bones. Differentiating chondrocytes in growth cartilage form four sequential zones of proliferation, alignment into column, hypertrophy, and substitution of chondrocytes with osteoblasts. Wnt/β-catenin signaling is essential for differentiation of proliferating chondrocytes into hypertrophic chondrocytes in growth cartilage. R-spondin 2 (Rspo2), a member of R-spondin family, is an agonist for Wnt signaling, but its role in chondrocyte differentiation remains unknown. Here we report that growth cartilage of Rspo2-knockout mice shows a decreased amount of β-catenin and increased amounts collagen type II (CII) and Sox9 in the abnormally extended proliferating zone. In contrast, expression of collagen type X (CX) in the hypertrophic zone remains unchanged. Differentiating chondrogenic ATDC5 cells, mimicking proliferating chondrocytes, upregulate Rspo2 and its putative receptor, Lgr5, in parallel. Addition of recombinant human Rspo2 to differentiating ATDC5 cells decreases expressions of Col2a1, Sox9, and Acan, as well as production of proteoglycans. In contrast, lentivirus-mediated knockdown of Rspo2 has the opposite effect. The effect of Rspo2 on chondrogenic differentiation is mediated by Wnt/β-catenin signaling, and not by Wnt/PCP or Wnt/Ca 2+ signaling. We propose that Rspo2 activates Wnt/β-catenin signaling to reduce Col2a1 and Sox9 and to facilitate differentiation of proliferating chondrocytes into hypertrophic chondrocytes in growth cartilage. - Highlights: • Rspo2 is a secreted activator of Wnt, and its knockout shows extended proliferating chondrocytes in endochondral ossification. • In proliferating chondrocytes of Rspo2-knockout mice, Sox9 and collagen type 2 are increased and β-catenin is decreased. • Rspo2 and its receptor Lgr5, as well as Sox9 and collagen type 2, are expressed in differentiating ATDC5 chondrogenic cells. • In ATDC5 cells, Rspo2 decreases expressions

  17. R-spondin 2 facilitates differentiation of proliferating chondrocytes into hypertrophic chondrocytes by enhancing Wnt/β-catenin signaling in endochondral ossification

    Energy Technology Data Exchange (ETDEWEB)

    Takegami, Yasuhiko [Division of Neurogenetics, Center of Neurological Disease and Cancer, Nagoya University Graduate School of Medicine, Nagoya (Japan); Department of Orthopaedic Surgery, Nagoya University School of Medicine, Nagoya (Japan); Ohkawara, Bisei; Ito, Mikako; Masuda, Akio [Division of Neurogenetics, Center of Neurological Disease and Cancer, Nagoya University Graduate School of Medicine, Nagoya (Japan); Nakashima, Hiroaki; Ishiguro, Naoki [Department of Orthopaedic Surgery, Nagoya University School of Medicine, Nagoya (Japan); Ohno, Kinji, E-mail: ohnok@med.nagoya-u.ac.jp [Division of Neurogenetics, Center of Neurological Disease and Cancer, Nagoya University Graduate School of Medicine, Nagoya (Japan)

    2016-04-22

    Endochondral ossification is a crucial process for longitudinal growth of bones. Differentiating chondrocytes in growth cartilage form four sequential zones of proliferation, alignment into column, hypertrophy, and substitution of chondrocytes with osteoblasts. Wnt/β-catenin signaling is essential for differentiation of proliferating chondrocytes into hypertrophic chondrocytes in growth cartilage. R-spondin 2 (Rspo2), a member of R-spondin family, is an agonist for Wnt signaling, but its role in chondrocyte differentiation remains unknown. Here we report that growth cartilage of Rspo2-knockout mice shows a decreased amount of β-catenin and increased amounts collagen type II (CII) and Sox9 in the abnormally extended proliferating zone. In contrast, expression of collagen type X (CX) in the hypertrophic zone remains unchanged. Differentiating chondrogenic ATDC5 cells, mimicking proliferating chondrocytes, upregulate Rspo2 and its putative receptor, Lgr5, in parallel. Addition of recombinant human Rspo2 to differentiating ATDC5 cells decreases expressions of Col2a1, Sox9, and Acan, as well as production of proteoglycans. In contrast, lentivirus-mediated knockdown of Rspo2 has the opposite effect. The effect of Rspo2 on chondrogenic differentiation is mediated by Wnt/β-catenin signaling, and not by Wnt/PCP or Wnt/Ca{sup 2+} signaling. We propose that Rspo2 activates Wnt/β-catenin signaling to reduce Col2a1 and Sox9 and to facilitate differentiation of proliferating chondrocytes into hypertrophic chondrocytes in growth cartilage. - Highlights: • Rspo2 is a secreted activator of Wnt, and its knockout shows extended proliferating chondrocytes in endochondral ossification. • In proliferating chondrocytes of Rspo2-knockout mice, Sox9 and collagen type 2 are increased and β-catenin is decreased. • Rspo2 and its receptor Lgr5, as well as Sox9 and collagen type 2, are expressed in differentiating ATDC5 chondrogenic cells. • In ATDC5 cells, Rspo2 decreases

  18. Detecting new microRNAs in human osteoarthritic chondrocytes identifies miR-3085 as a human, chondrocyte-selective, microRNA

    OpenAIRE

    Crowe, N.; Swingler, T.E.; Le, L.T.T.; Barter, M.J.; Wheeler, G.; Pais, H.; Donell, S.T.; Young, D.A.; Dalmay, T.; Clark, I.M.

    2016-01-01

    Summary Objective To use deep sequencing to identify novel microRNAs (miRNAs) in human osteoarthritic cartilage which have a functional role in chondrocyte phenotype or function. Design A small RNA library was prepared from human osteoarthritic primary chondrocytes using in-house adaptors and analysed by Illumina sequencing. Novel candidate miRNAs were validated by northern blot and qRT-PCR. Expression was measured in cartilage models. Targets of novel candidates were identified by microarray...

  19. Distant Galaxies, Black Holes and Other Celestial Phenomena: NASA's Chandra X-ray Observatory Marks Four Years of Discovery Firsts

    Science.gov (United States)

    2003-09-01

    . Building on previous achievements, including catching a supermassive black hole devouring material in our own Milky Way galaxy, Chandra accomplished even more during its fourth year. The observatory revealed new details about X-ray jets produced by black holes and discovered two black holes flourishing in a single galaxy 400 million light years from Earth. By tracking, for the first time, the life cycle of large-scale X-ray jets produced by a black hole, Chandra revealed that as the jets evolved, the material in them traveled near the speed of light for several years before slowing and fading. These jets - from a stellar-sized black hole about 10 or so times the mass of the Sun - were the first ones caught in the act of slowing down. This enabled astronomers, in just four years, to observe a process that could take a million years to unfold. By revealing two active black holes in the nucleus of the extraordinarily bright galaxy NGC 6240, another Chandra image proved for the first time that two supermassive black holes can co-exist in the same galaxy. Currently orbiting each other, in several hundred million years these black holes will merge to create an even larger black hole, resulting in a catastrophic event that will unleash intense radiation and gravitational waves. Also in Chandra's fourth year, the observatory offered new insights into pulsars - small and extremely dense stars. Generated by a series of Chandra observations, an X-ray movie of the Vela pulsar. revealed a spectacularly erratic jet that varied in a way never before seen. Whipping about like an untended firehose at about half the speed of light, the jet of high-energy particles offered new insight into the nature of jets from pulsars and black holes. Previous Chandra highlights include revealing the most distant X-ray cluster of galaxies, identifying a pulsating hot spot of X-rays in Jupiter's upper atmosphere, uncovering a ''cool'' black hole at the heart of the Andromeda Galaxy, and finding an X

  20. Cyclic Equibiaxial Tensile Strain Alters Gene Expression of Chondrocytes via Histone Deacetylase 4 Shuttling.

    Directory of Open Access Journals (Sweden)

    Chongwei Chen

    Full Text Available This paper aims to investigate whether equibiaxial tensile strain alters chondrocyte gene expression via controlling subcellular localization of histone deacetylase 4 (HDAC4.Murine chondrocytes transfected with GFP-HDAC4 were subjected to 3 h cyclic equibiaxial tensile strain (CTS, 6% strain at 0.25 Hz by a Flexcell® FX-5000™ Tension System. Fluorescence microscope and western blot were used to observe subcellular location of HDAC4. The gene expression was analyzed by real-time RT-PCR. The concentration of Glycosaminoglycans in culture medium was quantified by bimethylmethylene blue dye; Collagen II protein was evaluated by western blot. Cells phenotype was identified by immunohistochemistry. Cell viability was evaluated by live-dead cell detect kit. Okadaic acid, an inhibitor of HDAC4 nuclear relocation, was used to further validate whether HDAC4 nuclear relocation plays a role in gene expression in response to tension stimulation.87.5% of HDAC4 was located in the cytoplasm in chondrocytes under no loading condition, but it was relocated to the nucleus after CTS. RT-PCR analysis showed that levels of mRNA for aggrecan, collagen II, LK1 and SOX9 were all increased in chondrocytes subjected to CTS as compared to no loading control chondrocytes; in contrast, the levels of type X collagen, MMP-13, IHH and Runx2 gene expression were decreased in the chondrocytes subjected to CTS as compared to control chondrocytes. Meanwhile, CTS contributed to elevation of glycosaminoglycans and collagen II protein, but did not change collagen I production. When Okadaic acid blocked HDAC4 relocation from the cytoplasm to nucleus, the changes of the chondrocytes induced by CTS were abrogated. There was no chondrocyte dead detected in this study in response to CTS.CTS is able to induce HDAC4 relocation from cytoplasm to nucleus. Thus, CTS alters chondrocytes gene expression in association with the relocation of HDAC4 induced by CTS.

  1. Inflammation induction of Dickkopf-1 mediates chondrocyte apoptosis in osteoarthritic joint.

    Science.gov (United States)

    Weng, L-H; Wang, C-J; Ko, J-Y; Sun, Y-C; Su, Y-S; Wang, F-S

    2009-07-01

    Dysregulated Wnt signaling appears to modulate chondrocyte fate and joint disorders. Dickkopf-1 (DKK1) regulates the pathogenesis of skeletal tissue by inhibiting Wnt actions. This study examined whether DKK1 expression is linked to chondrocyte fate in osteoarthritis (OA). Articular cartilage specimens harvested from nine patients with knee OA and from six controls with femoral neck fracture were assessed for DKK1, interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), Bad, Bax, Bcl2 and caspase-3 expression by real time-polymerase chain reaction (RT-PCR) and immunohistochemistry. Apoptotic chondrocytes were detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labelling (TUNEL) and 4', 6-dianidino-2-phenylindole dihydrochloride (DAPI) staining. Human chondrocyte cultures were treated with recombinant IL-1beta and monoclonal DKK1 antibody to determine whether DKK1 impairs chondrocyte survival. Expression of DKK1 correlated with inflammatory cytokine levels (IL-1beta and TNF-alpha expressions), proapoptosis regulators (Bad and caspase-3 expressions) and TUNEL staining in OA cartilage tissues. The IL-1beta induced expressions of DKK1, Bax, Bad and caspase-3-dependent apoptosis of chondrocyte cultures. Neutralization of DKK1 by monoclonal DKK1 antibody significantly abrogated IL-1beta-mediated caspase-3 cleavage and apoptosis and reversed chondrocyte proliferation. Recombinant DKK1 treatment impaired chondrocyte growth and promoted apoptosis. By suppressing nuclear beta-catenin accumulation and Akt phosphorylation, DKK1 mediated IL-1beta promotion of chondrocyte apoptosis. Chondrocyte apoptosis correlates with joint OA. Expression of DKK1 contributes to cartilage deterioration and is a potent factor in OA pathogenesis. Attenuating DKK1 may reduce cartilage deterioration in OA.

  2. Four-year measurement of methane flux over a temperate forest with a relaxed eddy accumulation method

    Science.gov (United States)

    Sakabe, A.; Kosugi, Y.; Ueyama, M.; Hamotani, K.; Takahashi, K.; Iwata, H.; Itoh, M.

    2013-12-01

    Forests are generally assumed to be an atmospheric methane (CH4) sink (Le Mer and Roger, 2001). However, under Asian monsoon climate, forests are subject to wide spatiotemporal range in soil water status, where forest soils often became water-saturated condition heterogeneously. In such warm and humid conditions, forests may act as a CH4 source and/or sink with considerable spatiotemporal variations. Micrometeorological methods such as eddy covariance (EC) method continuously measure spatially-representative flux at a canopy scale without artificial disturbance. In this study, we measured CH4 fluxes over a temperate forest during four-year period using a CH4 analyzer based on tunable diode laser spectroscopy detection with a relaxed eddy accumulation (REA) method (Hamotani et al., 1996, 2001). We revealed the amplitude and seasonal variations of canopy-scale CH4 fluxes. The REA method is the attractive alternative to the EC method to measure trace-gas flux because it allows the use of analyzers with an optimal integration time. We also conducted continuous chamber measurements on forest floor to reveal spatial variations in soil CH4 fluxes and its controlling processes. The observations were made in an evergreen coniferous forest in central Japan. The site has a warm temperate monsoon climate with wet summer. Some wetlands were located in riparian zones along streams within the flux footprint area. For the REA method, the sonic anemometer (SAT-550, Kaijo) was mounted on top of the 29-m-tall tower and air was sampled from just below the sonic anemometer to reservoirs according to the direction of vertical wind velocity (w). After accumulating air for 30 minutes, the air in the reservoirs was pulled into a CO2/H2O gas analyzer (LI-840, Li-Cor) and a CH4 analyzer (FMA-200, Los Gatos Research). Before entering the analyzers, the sampled air was dried using a gas dryer (PD-50 T-48; Perma Pure Inc.). The REA flux is obtained from the difference in the mean concentrations

  3. A four-year experience with a Web-based self-help intervention for depressive symptoms in Mexico

    Directory of Open Access Journals (Sweden)

    María Asunción Lara

    2014-06-01

    Full Text Available OBJECTIVE: To describe a four-year descriptive, naturalistic study monitoring the use of HDep (Help for Depression or Ayuda para depression (ADepin Spanish, an open-access/free Web-based, psycho-education, cognitive-behavioral intervention program produced in Mexico consisting of seven self-help modules that include feedback-generating assessments of depressive symptoms, vignettes, recorded messages, a relaxation exercise, a personal workbook, blogs, and user discussion forums. METHODS: Data were collected on all individuals who entered the HDep site since the program's launching in 2009. Those who entered the site two or more times and also registered as "users" or "participants." The user data consisted of 1 user profiles; 2 scores for the CES-D (Center for Epidemiological Studies Depression Scale, for users who completed the feedback-generating assessments of depressive symptoms; 3 user evaluations of the usefulness of HDep; and 4 transcripts of HDep discussion forum posts. The raw user data were obtained through Moodle (Modular Object-Oriented Dynamic Learning Environment, a free software e-learning platform and analyzed quantitatively (using SPSS and qualitatively (using ATLAS.ti. RESULTS: A total of 28 078 individuals accessed HDep and 17 318 of those (61.6% qualified as users. Of all users, 84.4% were women, 64.6% used the workbook, and 60.9% entered the discussion forums (of whom 16.3% added a post. Depressive symptoms (CES-D score ≥ 16 were observed in 97.1% of the users who completed the feedback-generating assessment (n = 16 564. User retention dropped across the seven modules (from 12 366 users for Module 1 to 626 for Module 7. However, all seven modules were rated very high for "helpfulness/usefulness," with mean scores all above 4 on a 1 - 5 scale. The HDep discussion forums showed a rich social interaction. Predictors of entering at least one module (based on stepwise logistic regression analysis included being a woman, being

  4. Proteoglycon synthesis by articular chondrocytes in agarose culture

    International Nuclear Information System (INIS)

    Sweet, M.B.E.; Grisillo, A.; Coehlo, A.; Schnitzler, C.M.

    1987-01-01

    Articular chondrocytes were isolated from knee joints of full-term bovine foetuses and grown in long-term agarose cultures. At intervals, cultures were labelled with 35 S-[sulphate] or D[6- 3 H] glucosamine. Newly synthesized proteoglycans were extracted with 4 M guanidine HCl and purified by isopycnic density gradient centrifugation or on DEAE cellulose in the presence of 8 M urea. Characterization of the proteoglycans revealed them to be identical in size to those present in the tissue and to be similarly capable of aggregation with hyaluronate. Newly synthesized chondroitin sulphate chains were identical in size, but newly synthesized keratan sulphate chains were somewhat larger than those present in the tissue. The newly synthesized proteoglycans were shown to contain the same range of O-linked oligosaccharides identified in proteoglycans of the Swarm rat chondrosarcoma. Cartilage-specific proteoglycan continued to be synthesized by the chondrocytes for up to 60 days; however, with time, proportionately more of a small non-aggregating proteoglycan appeared

  5. The role of autologous chondrocyte implantation in the treatment of symptomatic chondromalacia patellae.

    Science.gov (United States)

    Macmull, Simon; Jaiswal, Parag K; Bentley, George; Skinner, John A; Carrington, Richard W J; Briggs, Tim W R

    2012-07-01

    Chondromalacia patella is a distinct clinical entity of abnormal softening of the articular cartilage of the patella, which results in chronic retropatellar pain. Its aetiology is still unclear but the process is thought to be a due to trauma to superficial chondrocytes resulting in a proteolytic enzymic breakdown of the matrix. Our aim was to assess the effectiveness of autologous chondrocyte implantation on patients with a proven symptomatic retropatellar lesion who had at least one failed conventional marrow-stimulating therapy. We performed chondrocyte implantation on 48 patients: 25 received autologous chondrocyte implantation with a type I/III membrane (ACI-C) method (Geistlich Biomaterials, Wolhusen, Switzerland), and 23 received the Matrix-assisted Chondrocyte Implantation (MACI) technique (Genzyme, Kastrup, Denmark). Over a mean follow-up period of 40.3 months, there was a statistically significant improvement in subjective pain scoring using the visual analogue scale (VAS) and objective functional scores using the Modified Cincinnati Rating System (MCS) in both groups. Chondromalacia patellae lesions responded well to chondrocyte implantation. Better results occurred with MACI than with ACI-C. Excellent and good results were achieved in 40% of ACI-C patients and 57% of MACI patients, but success of chondrocyte implantation was greater with medial/odd-facet lesions. Given that the MACI procedure is technically easier and less time consuming, we consider it to be useful for treating patients with symptomatic chondral defects secondary to chondromalacia patellae.

  6. Centrifugation assay for measuring adhesion of serially passaged bovine chondrocytes to polystyrene surfaces.

    Science.gov (United States)

    Kaplan, David S; Hitchins, Victoria M; Vegella, Thomas J; Malinauskas, Richard A; Ferlin, Kimberly M; Fisher, John P; Frondoza, Carmelita G

    2012-07-01

    A major obstacle in chondrocyte-based therapy for cartilage repair is the limited availability of cells that maintain their original phenotype. Propagation of chondrocytes as monolayer cultures on polystyrene surfaces is used extensively for amplifying cell numbers. However, chondrocytes undergo a phenotypic shift when propagated in this manner and display characteristics of more adherent fibroblastic cells. Little information is available about the effect of this phenotypic shift on cellular adhesion properties. We evaluated changes in adhesion property as bovine chondrocytes were serially propagated up to five passages in monolayer culture using a centrifugation cell adhesion assay, which was based on counting of cells before and after being exposed to centrifugal dislodgement forces of 120 and 350 g. Chondrocytes proliferated well in a monolayer culture with doubling times of 2-3 days, but they appeared more fibroblastic and exhibited elongated cell morphology with continued passage. The centrifugation cell adhesion assay showed that chondrocytes became more adhesive with passage as the percentage of adherent cells after centrifugation increased and was not statistically different from the adhesion of the fibroblast cell line, L929, starting at passage 3. This increased adhesiveness correlated with a shift to a fibroblastic morphology and increased collagen I mRNA expression starting at passage 2. Our findings indicate that the centrifugation cell adhesion assay may serve as a reproducible tool to track alterations in chondrocyte phenotype during their extended propagation in culture.

  7. Smad6/Smurf1 overexpression in cartilage delays chondrocyte hypertrophy and causes dwarfism with osteopenia

    Science.gov (United States)

    Horiki, Mitsuru; Imamura, Takeshi; Okamoto, Mina; Hayashi, Makoto; Murai, Junko; Myoui, Akira; Ochi, Takahiro; Miyazono, Kohei; Yoshikawa, Hideki; Tsumaki, Noriyuki

    2004-01-01

    Biochemical experiments have shown that Smad6 and Smad ubiquitin regulatory factor 1 (Smurf1) block the signal transduction of bone morphogenetic proteins (BMPs). However, their in vivo functions are largely unknown. Here, we generated transgenic mice overexpressing Smad6 in chondrocytes. Smad6 transgenic mice showed postnatal dwarfism with osteopenia and inhibition of Smad1/5/8 phosphorylation in chondrocytes. Endochondral ossification during development in these mice was associated with almost normal chondrocyte proliferation, significantly delayed chondrocyte hypertrophy, and thin trabecular bone. The reduced population of hypertrophic chondrocytes after birth seemed to be related to impaired bone growth and formation. Organ culture of cartilage rudiments showed that chondrocyte hypertrophy induced by BMP2 was inhibited in cartilage prepared from Smad6 transgenic mice. We then generated transgenic mice overexpressing Smurf1 in chondrocytes. Abnormalities were undetectable in Smurf1 transgenic mice. Mating Smad6 and Smurf1 transgenic mice produced double-transgenic pups with more delayed endochondral ossification than Smad6 transgenic mice. These results provided evidence that Smurf1 supports Smad6 function in vivo. PMID:15123739

  8. Chondrocyte survival in osteochondral transplant cylinders depends on the harvesting technique.

    Science.gov (United States)

    Hafke, Benedikt; Petri, Maximilian; Suero, Eduardo; Neunaber, Claudia; Kwisda, Sebastian; Krettek, Christian; Jagodzinski, Michael; Omar, Mohamed

    2016-07-01

    In autologous osteochondral transplantation, the edges of the harvested plug are particularly susceptible to mechanical or thermal damage to the chondrocytes. We hypothesised that the applied harvesting device has an impact on chondrocyte vitality. Both knees of five blackhead sheep (ten knees) underwent open osteochondral plug harvesting with three different circular harvesting devices (osteoarticular transfer system harvester [OATS; diameter 8 mm; Arthrex, Munich, Germany], diamond cutter [DC; diameter 8.35 mm; Karl Storz, Tuttlingen, Germany] and hollow reamer with cutting crown [HRCC; diameter 7 mm; Dannoritzer, Tuttlingen, Germany]) from distinctly assigned anatomical sites of the knee joint. The rotary cutters (DC and HRCC) were either used with (+) or without cooling (-). Surgical cuts of the cartilage with a scalpel blade were chosen as control method. After cryotomy cutting, chondrocyte vitality was assessed using fluorescence microscopy and a Live/Dead assay. There were distinct patterns of chondrocyte vitality, with reproducible accumulations of dead chondrocytes along the harvesting edge. No statistical difference in chondrocyte survivorship was seen between the OATS technique and the control method, or between the HRCC+ technique and the control method (P > 0.05). The DC+, HRCC- and DC- techniques yielded significantly lower chondrocyte survival rates compared with the control method (P vitality.

  9. Culture temperature affects redifferentiation and cartilaginous extracellular matrix formation in dedifferentiated human chondrocytes.

    Science.gov (United States)

    Ito, Akira; Aoyama, Tomoki; Iijima, Hirotaka; Tajino, Junichi; Nagai, Momoko; Yamaguchi, Shoki; Zhang, Xiangkai; Kuroki, Hiroshi

    2015-05-01

    To date, there have been few studies on how temperature affects the phenotype and metabolism of human chondrocytes. Thus, the purpose of this study was to elucidate the effects of culture temperature on chondrocyte redifferentiation and extracellular matrix (ECM) formation using dedifferentiated mature human chondrocytes in vitro. Dedifferentiated chondrocytes were cultured in a pellet culture system for up to 21 days. The pellets were randomly divided into three groups with different culture temperature (32, 37, and 41°C). Chondrocyte redifferentiation and ECM formation were evaluated by wet weight, messenger ribonucleic acid (mRNA), histological, and biochemical analyses. The results showed that the wet weight and the mRNA expressions of collagen type II A1 and cartilage oligomeric matrix protein at 37°C were higher than the corresponding values at 32°C. The histological and biochemical analyses revealed that the syntheses of type II collagen and proteoglycan were promoted at 37°C compared to those at 32°C, whereas they were considerably inhibited at 41°C. In conclusion, the results obtained herein indicated that temperature affects chondrocyte redifferentiation and ECM formation, and modulation of temperature might thus represent an advantageous means to regulate the phenotype and biosynthetic activity of chondrocytes. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  10. Chondrocyte differentiation for auricular cartilage reconstruction using a chitosan based hydrogel.

    Science.gov (United States)

    García-López, J; Garciadiego-Cázares, D; Melgarejo-Ramírez, Y; Sánchez-Sánchez, R; Solís-Arrieta, L; García-Carvajal, Z; Sánchez-Betancourt, J I; Ibarra, C; Luna-Bárcenas, G; Velasquillo, C

    2015-12-01

    Tissue engineering with the use of biodegradable and biocompatible scaffolds is an interesting option for ear repair. Chitosan-Polyvinyl alcohol-Epichlorohydrine hydrogel (CS-PVA-ECH) is biocompatible and displays appropriate mechanical properties to be used as a scaffold. The present work, studies the potential of CS-PVA-ECH scaffolds seeded with chondrocytes to develop elastic cartilage engineered-neotissues. Chondrocytes isolated from rabbit and swine elastic cartilage were independently cultured onto CS-PVA-ECH scaffolds for 20 days to form the appropriate constructs. Then, in vitro cell viability and morphology were evaluated by calcein AM and EthD-1 assays and Scanning Electron Microscopy (SEM) respectively, and the constructs were implanted in nu/nu mice for four months, in order to evaluate the neotissue formation. Histological analysis of the formed neotissues was performed by Safranin O, Toluidine blue (GAG's), Verhoeff-Van Gieson (elastic fibers), Masson's trichrome (collagen) and Von Kossa (Calcium salts) stains and SEM. Results indicate appropriate cell viability, seeded with rabbit or swine chondrocyte constructs; nevertheless, upon implantation the constructs developed neotissues with different characteristics depending on the animal species from which the seeded chondrocytes came from. Neotissues developed from swine chondrocytes were similar to auricular cartilage, while neotissues from rabbit chondrocytes were similar to hyaline cartilage and eventually they differentiate to bone. This result suggests that neotissue characteristics may be influenced by the animal species source of the chondrocytes isolated.

  11. Effects of intermittent versus continuous parathyroid hormone administration on condylar chondrocyte proliferation and differentiation

    International Nuclear Information System (INIS)

    Liu, Qi; Wan, Qilong; Yang, Rongtao; Zhou, Haihua; Li, Zubing

    2012-01-01

    Highlights: ► Different PTH administration exerts different effects on condylar chondrocyte. ► Intermittent PTH administration suppresses condylar chondrocyte proliferation. ► Continuous PTH administration maintains condylar chondrocyte proliferating. ► Intermittent PTH administration enhances condylar chondrocyte differentiation. -- Abstract: Endochondral ossification is a complex process involving chondrogenesis and osteogenesis regulated by many hormones and growth factors. Parathyroid hormone (PTH), one of the key hormones regulating bone metabolism, promotes osteoblast differentiation and osteogenesis by intermittent administration, whereas continuous PTH administration inhibits bone formation. However, the effects of PTH on chondrocyte proliferation and differentiation are still unclear. In this study, intermittent PTH administration presented enhanced effects on condylar chondrocyte differentiation and bone formation, as demonstrated by increased mineral nodule formation and alkaline phosphatase (ALP) activity, up-regulated runt-related transcription factor 2 (RUNX2), ALP, collagen type X (COL10a1), collagen type I (COL1a1), osteocalcin (OCN), bone sialoprotein (BSP), bone morphogenetic protein 2 (BMP2) and osterix (OSX) mRNA and/or protein expression. On the contrary, continuous PTH administration promoted condylar chondrocyte proliferation and suppressed its differentiation, as demonstrated by up-regulated collagen type II (COL2a1) mRNA expression, reduced mineral nodule formation and down-regulated expression of the mRNAs and/or proteins mentioned above. Our data suggest that PTH can regulate condylar chondrocyte proliferation and differentiation, depending on the type of PTH administration. These results provide new insight into the effects of PTH on condylar chondrocytes and new evidence for using local PTH administration to cure mandibular asymmetry.

  12. Effects of intermittent versus continuous parathyroid hormone administration on condylar chondrocyte proliferation and differentiation

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Qi; Wan, Qilong; Yang, Rongtao; Zhou, Haihua [The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan 430079 (China); Li, Zubing, E-mail: lizubing0827@163.com [The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan 430079 (China); Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan 430079 (China)

    2012-07-20

    Highlights: Black-Right-Pointing-Pointer Different PTH administration exerts different effects on condylar chondrocyte. Black-Right-Pointing-Pointer Intermittent PTH administration suppresses condylar chondrocyte proliferation. Black-Right-Pointing-Pointer Continuous PTH administration maintains condylar chondrocyte proliferating. Black-Right-Pointing-Pointer Intermittent PTH administration enhances condylar chondrocyte differentiation. -- Abstract: Endochondral ossification is a complex process involving chondrogenesis and osteogenesis regulated by many hormones and growth factors. Parathyroid hormone (PTH), one of the key hormones regulating bone metabolism, promotes osteoblast differentiation and osteogenesis by intermittent administration, whereas continuous PTH administration inhibits bone formation. However, the effects of PTH on chondrocyte proliferation and differentiation are still unclear. In this study, intermittent PTH administration presented enhanced effects on condylar chondrocyte differentiation and bone formation, as demonstrated by increased mineral nodule formation and alkaline phosphatase (ALP) activity, up-regulated runt-related transcription factor 2 (RUNX2), ALP, collagen type X (COL10a1), collagen type I (COL1a1), osteocalcin (OCN), bone sialoprotein (BSP), bone morphogenetic protein 2 (BMP2) and osterix (OSX) mRNA and/or protein expression. On the contrary, continuous PTH administration promoted condylar chondrocyte proliferation and suppressed its differentiation, as demonstrated by up-regulated collagen type II (COL2a1) mRNA expression, reduced mineral nodule formation and down-regulated expression of the mRNAs and/or proteins mentioned above. Our data suggest that PTH can regulate condylar chondrocyte proliferation and differentiation, depending on the type of PTH administration. These results provide new insight into the effects of PTH on condylar chondrocytes and new evidence for using local PTH administration to cure mandibular

  13. Nuclear deformation and expression change of cartilaginous genes during in vitro expansion of chondrocytes

    International Nuclear Information System (INIS)

    Hoshiba, Takashi; Yamada, Tomoe; Lu, Hongxu; Kawazoe, Naoki; Tateishi, Tetsuya; Chen, Guoping

    2008-01-01

    Cartilaginous gene expression decreased when chondrocytes were expanded on cell-culture plates. Understanding the dedifferentiation mechanism may provide valuable insight into cartilage tissue engineering. Here, we demonstrated the relationship between the nuclear shape and gene expression during in vitro expansion culture of chondrocytes. Specifically, the projected nuclear area increased and cartilaginous gene expressions decreased during in vitro expansion culture. When the nuclear deformation was recovered by cytochalasin D treatment, aggrecan expression was up-regulated and type I collagen (Col1a2) expression was down-regulated. These results suggest that nuclear deformation may be one of the mechanisms for chondrocyte dedifferentiation during in vitro expansion culture

  14. Graft-Sparing Strategy for Thoracic Prosthetic Graft Infection.

    Science.gov (United States)

    Uchino, Gaku; Yoshida, Takeshi; Kakii, Bunpachi; Furui, Masato

    2018-04-01

     Thoracic prosthetic graft infection is a rare but serious complication with no standard management. We reported our surgical experience on graft-sparing strategy for thoracic prosthetic graft infection.  This study included patients who underwent graft-sparing surgery for thoracic prosthetic graft infection at Matsubara Tokushukai Hospital in Japan from January 2000 to October 2017.  There were 17 patients included in the analyses, with a mean age at surgery of 71.0 ± 10.5 years; 11 were men. In-hospital mortality was observed in five patients (29.4%).  Graft-sparing surgery for thoracic prosthetic graft infection is an alternative option particularly for early graft infection after hemiarch replacement. Georg Thieme Verlag KG Stuttgart · New York.

  15. Rapid Chondrocyte Isolation for Tissue Engineering Applications: The Effect of Enzyme Concentration and Temporal Exposure on the Matrix Forming Capacity of Nasal Derived Chondrocytes

    Directory of Open Access Journals (Sweden)

    Srujana Vedicherla

    2017-01-01

    Full Text Available Laboratory based processing and expansion to yield adequate cell numbers had been the standard in Autologous Disc Chondrocyte Transplantation (ADCT, Allogeneic Juvenile Chondrocyte Implantation (NuQu®, and Matrix-Induced Autologous Chondrocyte Implantation (MACI. Optimizing cell isolation is a key challenge in terms of obtaining adequate cell numbers while maintaining a vibrant cell population capable of subsequent proliferation and matrix elaboration. However, typical cell yields from a cartilage digest are highly variable between donors and based on user competency. The overall objective of this study was to optimize chondrocyte isolation from cartilaginous nasal tissue through modulation of enzyme concentration exposure (750 and 3000 U/ml and incubation time (1 and 12 h, combined with physical agitation cycles, and to assess subsequent cell viability and matrix forming capacity. Overall, increasing enzyme exposure time was found to be more detrimental than collagenase concentration for subsequent viability, proliferation, and matrix forming capacity (sGAG and collagen of these cells resulting in nonuniform cartilaginous matrix deposition. Taken together, consolidating a 3000 U/ml collagenase digest of 1 h at a ratio of 10 ml/g of cartilage tissue with physical agitation cycles can improve efficiency of chondrocyte isolation, yielding robust, more uniform matrix formation.

  16. Hyperpolarisation of cultured human chondrocytes following cyclical pressure-induced strain: evidence of a role for alpha 5 beta 1 integrin as a chondrocyte mechanoreceptor.

    Science.gov (United States)

    Wright, M O; Nishida, K; Bavington, C; Godolphin, J L; Dunne, E; Walmsley, S; Jobanputra, P; Nuki, G; Salter, D M

    1997-09-01

    Mechanical stimuli influence chondrocyte metabolism, inducing changes in intracellular cyclic adenosine monophosphate and proteoglycan production. We have previously demonstrated that primary monolayer cultures of human chondrocytes have an electrophysiological response after intermittent pressure-induced strain characterised by a membrane hyperpolarisation of approximately 40%. The mechanisms responsible for these changes are not fully understood but potentially involve signalling molecules such as integrins that link extracellular matrix with cytoplasmic components. The results reported in this paper demonstrate that the transduction pathways involved in the hyperpolarisation response of human articular chondrocytes in vitro after cyclical pressure-induced strain involve alpha 5 beta 1 integrin. We have demonstrated, using pharmacological inhibitors of a variety of intracellular signalling pathways, that the actin cytoskeleton, the phospholipase C calmodulin pathway, and both tyrosine protein kinase and protein kinase C activities are important in the transduction of the electrophysiological response. These results suggest that alpha 5 beta 1 is an important chondrocyte mechanoreceptor and a potential regulator of chondrocyte function.

  17. Growth factor effects on costal chondrocytes for tissue engineering fibrocartilage

    Science.gov (United States)

    Johns, D.E.; Athanasiou, K.A.

    2010-01-01

    Tissue engineered fibrocartilage could become a feasible option for replacing tissues like the knee meniscus or temporomandibular joint disc. This study employed five growth factors insulin-like growth factor-I, transforming growth factor-β1, epidermal growth factor, platelet-derived growth factor-BB, and basic fibroblast growth factor in a scaffoldless approach with costal chondrocytes, attempting to improve biochemical and mechanical properties of engineered constructs. Samples were quantitatively assessed for total collagen, glycosaminoglycans, collagen type I, collagen type II, cells, compressive properties, and tensile properties at two time points. Most treated constructs were worse than the no growth factor control, suggesting a detrimental effect, but the IGF treatment tended to improve the constructs. Additionally, the 6wk time point was consistently better than 3wks, with total collagen, glycosaminoglycans, and aggregate modulus doubling during this time. Further optimization of the time in culture and exogenous stimuli will be important in making a more functional replacement tissue. PMID:18597118

  18. Is gender influencing the biomechanical results after autologous chondrocyte implantation?

    Science.gov (United States)

    Kreuz, Peter C; Müller, Sebastian; Erggelet, Christoph; von Keudell, Arvind; Tischer, Thomas; Kaps, Christian; Niemeyer, Philipp; Hirschmüller, Anja

    2014-01-01

    The influence of gender on the biomechanical outcome after autologous chondrocyte implantation (ACI) including isokinetic muscle strength measurements has not been investigated. The present prospective study was performed to evaluate gender-specific differences in the biomechanical function 48 months after ACI. Fifty-two patients (mean age 35.6 ± 8.5 years) that met our inclusion criteria, underwent ACI with Bioseed C(®) and were evaluated with the KOOS score preoperatively, 6, 12 and 48 months after surgery. At final follow-up, 44 out of the 52 patients underwent biomechanical evaluation with isokinetic strength measurements of both knees. All data were evaluated separately for men and women and compared for each time interval using the Mann-Whitney U test. Clinical scores improved significantly over the whole study period (p genders. Isokinetic muscle strength measures are significantly worse in women (p role for the explanation of gender-specific results after ACI.

  19. Streptococcus pyogenes degrades extracellular matrix in chondrocytes via MMP-13

    International Nuclear Information System (INIS)

    Sakurai, Atsuo; Okahashi, Nobuo; Maruyama, Fumito; Ooshima, Takashi; Hamada, Shigeyuki; Nakagawa, Ichiro

    2008-01-01

    Group A streptococcus (GAS) causes a wide range of human diseases, including bacterial arthritis. The pathogenesis of arthritis is characterized by synovial proliferation and the destruction of cartilage and subchondral bone in joints. We report here that GAS strain JRS4 invaded a chondrogenic cell line ATDC5 and induced the degradation of the extracellular matrix (ECM), whereas an isogenic mutant of JRS4 lacking a fibronectin-binding protein, SAM1, failed to invade the chondrocytes or degrade the ECM. Reverse transcription-PCR and Western blot analysis revealed that the expression of matrix metalloproteinase (MMP)-13 was strongly elevated during the infection with GAS. A reporter assay revealed that the activation of the AP-1 transcription factor and the phosphorylation of c-Jun terminal kinase participated in MMP-13 expression. These results suggest that MMP-13 plays an important role in the destruction of infected joints during the development of septic arthritis

  20. Auditory-Visual Speech Perception in Three- and Four-Year-Olds and Its Relationship to Perceptual Attunement and Receptive Vocabulary

    Science.gov (United States)

    Erdener, Dogu; Burnham, Denis

    2018-01-01

    Despite the body of research on auditory-visual speech perception in infants and schoolchildren, development in the early childhood period remains relatively uncharted. In this study, English-speaking children between three and four years of age were investigated for: (i) the development of visual speech perception--lip-reading and visual…

  1. Sustaining the Support in Four-Year-Olds in Childcare Services with the Goal of Promoting Their Cognitive and Language Development

    Science.gov (United States)

    Bigras, Nathalie; Lemay, Lise; Bouchard, Caroline; Eryasa, Joell

    2017-01-01

    This paper aims to describe the quality of play as offered by early childhood educators working with four-year-olds within an educational childcare service. It also aims to identify the correlation between the quality of play support and a child's cognitive, language and socio-emotional development. Finally, it focuses on the factors associated…

  2. Designing for Diverse Classrooms: Using iPpads and Digital Cameras to Compose eBooks with Emergent Bilingual/Biliterate Four-Year-Olds

    Science.gov (United States)

    Rowe, Deborah Wells; Miller, Mary E.

    2016-01-01

    This paper reports the findings of a two-year design study exploring instructional conditions supporting emerging, bilingual/biliterate, four-year-olds' digital composing. With adult support, children used child-friendly, digital cameras and iPads equipped with writing, drawing and bookmaking apps to compose multimodal, multilingual eBooks…

  3. Simultaneous vs. Successive Bilingualism among Preschool-Aged Children: A Study of Four-Year-Old Korean-English Bilinguals in the USA

    Science.gov (United States)

    Kim, Ah-Young; Park, Anne; Lust, Barbara

    2018-01-01

    The current study compares simultaneous vs. successive bilingualism through a mixed-method research design with four four-year-old Korean-English bilingual children who were born and raised in the USA. Two simultaneous bilinguals were exposed to Korean and English from birth, whereas two successive bilinguals were exposed to Korean from birth, but…

  4. Arteriovenous shunt graft ulceration with sinus and graft epithelialization

    Directory of Open Access Journals (Sweden)

    Pooja Singhal

    2015-03-01

    Full Text Available Arteriovenous fistula and grafts are used as access sites for patients with chronic kidney disease and are prone for complications. Stent grafts are used to treat access site complications. We report a rare and unusual finding of epithelialization of the sinus tract and the lumen of a polytetrafluoroethylene graft, following ulceration of the overlying skin.

  5. Chitosan-Graft-Polyethylenimine/DNA Nanoparticles as Novel Non-Viral Gene Delivery Vectors Targeting Osteoarthritis

    Science.gov (United States)

    Lv, Lulu; Zhao, Huiqing

    2014-01-01

    The development of safe and efficient gene carriers is the key to the clinical success of gene therapy. The present study was designed to develop and evaluate the chitosan-graft-polyethylenimine (CP)/DNA nanoparticles as novel non-viral gene vectors for gene therapy of osteoarthritis. The CP/DNA nanoparticles were produced through a complex coacervation of the cationic polymers with pEGFP after grafting chitosan (CS) with a low molecular weight (Mw) PEI (Mw = 1.8 kDa). Particle size and zeta potential were related to the weight ratio of CP:DNA, where decreases in nanoparticle size and increases in surface charge were observed as CP content increased. The buffering capacity of CP was significantly greater than that of CS. The transfection efficiency of CP/DNA nanoparticles was similar with that of the Lipofectamine™ 2000, and significantly higher than that of CS/DNA and PEI (25 kDa)/DNA nanoparticles. The transfection efficiency of the CP/DNA nanoparticles was dependent on the weight ratio of CP:DNA (w/w). The average cell viability after the treatment with CP/DNA nanoparticles was over 90% in both chondrocytes and synoviocytes, which was much higher than that of PEI (25 kDa)/DNA nanoparticles. The CP copolymers efficiently carried the pDNA inside chondrocytes and synoviocytes, and the pDNA was detected entering into nucleus. These results suggest that CP/DNA nanoparticles with improved transfection efficiency and low cytotoxicity might be a safe and efficient non-viral vector for gene delivery to both chondrocytes and synoviocytes. PMID:24392152

  6. A Longitudinal Study on Substance Use and Related Problems in Women in Opioid Maintenance Treatment from Pregnancy to Four Years after Giving Birth

    Directory of Open Access Journals (Sweden)

    Ingunn O. Lund

    2014-01-01

    Full Text Available Background Women in opioid maintenance treatment (OMT have a past characterized by drug abuse, which is a challenging start for parenthood. Studies of mothers in OMT are typically limited to pregnancy and early infancy. Knowledge about how they cope with substance use and related problems in the years following birth is therefore important. The aims of the study were to examine changes in mothers’ substance use, psychological problems, and other challenges; from one to four years after their children were born, and describe kindergarten attendance and prevalence and type of child protective services involvement when the children were four years old. Method A four-year prospective cohort study of mothers in OMT. The European severity index was used to map substance use and related problems during the third trimester of pregnancy, one and four years after birth. Results At the four-year follow-up, use of illegal substances remained low (4% and use of legal substances (39% was similar to the one-year follow-up. The proportion of women with psychological problems was significantly higher than at one-year follow-up (69 vs. 39%, P = .009. At age four, most children (89% attended kindergarten, and the child protective services were following 73% of the families, mostly with voluntary measures. Conclusion Mothers in OMT cope well with substance use over time, given access to sufficient support. The findings imply that a preventive governmental strategy with close support of mother and child, have a positive impact contributing to making OMT and motherhood more compatible.

  7. The Regulatory Role of Signaling Crosstalk in Hypertrophy of MSCs and Human Articular Chondrocytes

    NARCIS (Netherlands)

    Zhong, Leilei; Huang, X; Karperien, Hermanus Bernardus Johannes; Post, Janine Nicole

    2015-01-01

    Hypertrophic differentiation of chondrocytes is a main barrier in application of mesenchymal stem cells (MSCs) for cartilage repair. In addition, hypertrophy occurs occasionally in osteoarthritis (OA). Here we provide a comprehensive review on recent literature describing signal pathways in the

  8. Effects of scaffold composition and architecture on human nasal chondrocyte redifferentiation and cartilaginous matrix deposition

    NARCIS (Netherlands)

    Miot, Sylvie; Woodfield, T.B.F.; Daniels, Alma U.; Suetterlin, Rosemarie; Peterschmitt, Iman; Heberer, Michael; van Blitterswijk, Clemens; Riesle, J.U.; Martin, Ivan

    2005-01-01

    We investigated whether the post-expansion redifferentiation and cartilage tissue formation capacity of adult human nasal chondrocytes can be regulated by controlled modifications of scaffold composition and architecture. As a model system, we used poly(ethylene

  9. Synthesis of collagen by bovine chondrocytes cultured in alginate; posttranslational modifications and cell-matrix interaction

    NARCIS (Netherlands)

    Beekman, B.; Verzijl, N.; Bank, R.A.; Von Der Mark, K.; TeKoppele, J.M.

    1997-01-01

    The extracellular matrix synthesized by articular chondrocytes cultured in alginate beads was investigated. Collagen levels increased sigmoidally with time and remained constant after 2 weeks of culture. The presence of cartilage-specific type II collagen was confirmed immunohistochemically.

  10. A Preliminary Study of Human Amniotic Membrane as a Potential Chondrocyte Carrier

    Directory of Open Access Journals (Sweden)

    L Boo

    2009-11-01

    Full Text Available PURPOSE: To investigate the feasibility of using processed human amniotic membrane (HAM to support the attachment and proliferation of chondrocytes in vitro which in turn can be utilised as a cell delivery vehicle in tissue engineering applications. METHODS: Fresh HAM obtained from patients undergoing routine elective caesarean sections was harvested, processed and dried using either freeze drying (FD or air drying (AD methods prior to sterilisation by gamma irradiation. Isolated, processed and characterised rabbit autologous chondrocytes were seeded on processed HAM and cultured for up to three weeks. Cell attachment and proliferation were examined qualitatively using inverted brightfield microscopy. RESULTS: Processed HAM appeared to allow cell attachment when implanted with chondrocytes. Although cells seeded on AD and FD HAM did not appear to attach as strongly as those seeded on glycerol preserved intact human amniotic membrane, these cells to be proliferated in cell culture conditions. CONCLUSION: Preliminary results show that processed HAM promotes chondrocyte attachment and proliferation.

  11. Beta1 integrins regulate chondrocyte rotation, G1 progression, and cytokinesis

    DEFF Research Database (Denmark)

    Aszodi, Attila; Hunziker, Ernst B; Brakebusch, Cord

    2003-01-01

    Beta1 integrins are highly expressed on chondrocytes, where they mediate adhesion to cartilage matrix proteins. To assess the functions of beta1 integrin during skeletogenesis, we inactivated the beta1 integrin gene in chondrocytes. We show here that these mutant mice develop a chondrodysplasia...... of various severity. beta1-deficient chondrocytes had an abnormal shape and failed to arrange into columns in the growth plate. This is caused by a lack of motility, which is in turn caused by a loss of adhesion to collagen type II, reduced binding to and impaired spreading on fibronectin, and an abnormal F......-actin organization. In addition, mutant chondrocytes show decreased proliferation caused by a defect in G1/S transition and cytokinesis. The G1/S defect is, at least partially, caused by overexpression of Fgfr3, nuclear translocation of Stat1/Stat5a, and up-regulation of the cell cycle inhibitors p16 and p21...

  12. ADAM12-S stimulates bone growth in transgenic mice by modulating chondrocyte proliferation and maturation

    DEFF Research Database (Denmark)

    Kveiborg, Marie; Albrechtsen, Reidar; Rudkjaer, Lise

    2006-01-01

    ADAM12-S transgenic mice exhibit a pronounced increase in the length of bones, such as femur, tibia, and vertebrae. The effect of ADAM12-S on longitudinal bone growth involves the modulation of chondrocyte proliferation and maturation, likely through proteolytic activities and altered cell......: Transgenic mice expressing the secreted form of human ADAM12, ADAM12-S, or a truncated metalloprotease-deficient form of ADAM12-S in the circulation were used to study the effects of ADAM12 on the skeleton. In addition, murine chondrocyte cultures were used to study the effect of ADAM12-S on cell...... studies showed that ADAM12-S inhibits chondrocyte adhesion to fibronectin and collagen type II. CONCLUSIONS: ADAM12-S stimulates bone growth in mice by modulating chondrocyte proliferation and maturation through mechanisms probably involving both metalloprotease and adhesion activities....

  13. Exploration of mechanisms underlying the strain-rate-dependent mechanical property of single chondrocytes

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, Trung Dung; Gu, YuanTong, E-mail: yuantong.gu@qut.edu.au [School of Chemistry, Physics and Mechanical Engineering, Queensland University of Technology, Brisbane, Queensland (Australia)

    2014-05-05

    Based on the characterization by Atomic Force Microscopy, we report that the mechanical property of single chondrocytes has dependency on the strain-rates. By comparing the mechanical deformation responses and the Young's moduli of living and fixed chondrocytes at four different strain-rates, we explore the deformation mechanisms underlying this dependency property. We found that the strain-rate-dependent mechanical property of living cells is governed by both of the cellular cytoskeleton and the intracellular fluid when the fixed chondrocytes are mainly governed by their intracellular fluid, which is called the consolidation-dependent deformation behavior. Finally, we report that the porohyperelastic constitutive material model which can capture the consolidation-dependent behavior of both living and fixed chondrocytes is a potential candidature to study living cell biomechanics.

  14. Second-generation autologous chondrocyte transplantation: MRI findings and clinical correlations at a minimum 5-year follow-up

    International Nuclear Information System (INIS)

    Kon, E.; Di Martino, A.; Filardo, G.; Tetta, C.; Busacca, M.; Iacono, F.; Delcogliano, M.; Albisinni, U.; Marcacci, M.

    2011-01-01

    Objective: To evaluate the clinical outcome of hyaluronan-based arthroscopic autologous chondrocyte transplantation at a minimum of 5 years of follow-up and to correlate it with the MRI evaluation parameters. Methods: Fifty consecutive patients were included in the study and evaluated clinically using the Cartilage Standard Evaluation Form as proposed by ICRS and the Tegner score. Forty lesions underwent MRI evaluation at a minimum 5-year follow-up. For the description and evaluation of the graft, we employed the MOCART-scoring system. Results: A statistically significant improvement in all clinical scores was observed at 2 and over 5 years. The total MOCART score and the signal intensity (3D-GE-FS) of the repair tissue were statistically correlated to the IKDC subjective evaluation. Larger size of the treated cartilage lesions had a negative influence on the degree of defect repair and filling, the integration to the border zone and the subchondral lamina integrity, whereas more intensive sport activity had a positive influence on the signal intensity of the repair tissue, the repair tissue surface, and the clinical outcome. Conclusion: Our findings confirm the durability of the clinical results obtained with Hyalograft C and the usefulness of MRI as a non-invasive method for the evaluation of the repaired tissue and the outcome after second-generation autologous transplantation over time.

  15. Second-generation autologous chondrocyte transplantation: MRI findings and clinical correlations at a minimum 5-year follow-up

    Energy Technology Data Exchange (ETDEWEB)

    Kon, E. [Biomechanics Laboratory, III Clinic, Rizzoli Orthopaedic Institute, Via Di Barbiano 1/10, 40136 Bologna (Italy); Di Martino, A., E-mail: a.dimartino@biomec.ior.it [Biomechanics Laboratory, III Clinic, Rizzoli Orthopaedic Institute, Via Di Barbiano 1/10, 40136 Bologna (Italy); Filardo, G. [Biomechanics Laboratory, III Clinic, Rizzoli Orthopaedic Institute, Via Di Barbiano 1/10, 40136 Bologna (Italy); Tetta, C.; Busacca, M. [Radiology, Rizzoli Orthopaedic Institute, Bologna (Italy); Iacono, F. [Biomechanics Laboratory, III Clinic, Rizzoli Orthopaedic Institute, Via Di Barbiano 1/10, 40136 Bologna (Italy); Delcogliano, M. [Orthopaedic Departement San Carlo di Nancy Hospital, Rome (Italy); Albisinni, U. [Radiology, Rizzoli Orthopaedic Institute, Bologna (Italy); Marcacci, M. [Biomechanics Laboratory, III Clinic, Rizzoli Orthopaedic Institute, Via Di Barbiano 1/10, 40136 Bologna (Italy)

    2011-09-15

    Objective: To evaluate the clinical outcome of hyaluronan-based arthroscopic autologous chondrocyte transplantation at a minimum of 5 years of follow-up and to correlate it with the MRI evaluation parameters. Methods: Fifty consecutive patients were included in the study and evaluated clinically using the Cartilage Standard Evaluation Form as proposed by ICRS and the Tegner score. Forty lesions underwent MRI evaluation at a minimum 5-year follow-up. For the description and evaluation of the graft, we employed the MOCART-scoring system. Results: A statistically significant improvement in all clinical scores was observed at 2 and over 5 years. The total MOCART score and the signal intensity (3D-GE-FS) of the repair tissue were statistically correlated to the IKDC subjective evaluation. Larger size of the treated cartilage lesions had a negative influence on the degree of defect repair and filling, the integration to the border zone and the subchondral lamina integrity, whereas more intensive sport activity had a positive influence on the signal intensity of the repair tissue, the repair tissue surface, and the clinical outcome. Conclusion: Our findings confirm the durability of the clinical results obtained with Hyalograft C and the usefulness of MRI as a non-invasive method for the evaluation of the repaired tissue and the outcome after second-generation autologous transplantation over time.

  16. Hyaluronan Protects Bovine Articular Chondrocytes against Cell Death Induced by Bupivacaine under Supraphysiologic Temperatures

    Science.gov (United States)

    Liu, Sen; Zhang, Qing-Song; Hester, William; O’Brien, Michael J.; Savoie, Felix H.; You, Zongbing

    2013-01-01

    Background Bupivacaine and supraphysiologic temperature can independently reduce cell viability of articular chondrocytes. In combination these two deleterious factors could further impair cell viability. Hypothesis Hyaluronan may protect chondrocytes from death induced by bupivacaine at supraphysiologic temperatures. Study Design Controlled laboratory study. Methods Bovine articular chondrocytes were treated with hyaluronan at physiologic (37°C) and supraphysiologic temperatures (45°C and 50°C) for one hour, and then exposed to bupivacaine for one hour at room temperature. Cell viability was assessed at three time points: immediately after treatment, six hours later, and twenty-four hours later using flow cytometry and fluorescence microscopy. The effects of hyaluronan on the levels of sulfated glycosaminoglycan in the chondrocytes were determined using Alcian blue staining. Results (1) Bupivacaine alone did not induce noticeable chondrocyte death at 37°C; (2) bupivacaine and temperature synergistically increased chondrocyte death, that is, when the chondrocytes were conditioned to 45°C and 50°C, 0.25% and 0.5% bupivacaine increased the cell death rate by 131% to 383% in comparison to the phosphate-buffered saline control group; and, (3) addition of hyaluronan reduced chondrocyte death rates to approximately 14% and 25% at 45°C and 50°C, respectively. Hyaluronan’s protective effects were still observed at six and twenty-four hours after bupivacaine treatment at 45°C. However, at 50°C, hyaluronan delayed but did not prevent the cell death caused by bupivacaine. One-hour treatment with hyaluronan significantly increased sulfated glycosaminoglycan levels in the chondrocytes. Conclusions Bupivacaine and supraphysiologic temperature synergistically increase chondrocyte death and hyaluronan may protect articular chondrocytes from death caused by bupivacaine. Clinical Relevance This study provides a rationale to perform pre-clinical and clinical studies to

  17. Effect of microcavitary alginate hydrogel with different pore sizes on chondrocyte culture for cartilage tissue engineering

    International Nuclear Information System (INIS)

    Zeng, Lei; Yao, Yongchang; Wang, Dong-an; Chen, Xiaofeng

    2014-01-01

    In our previous work, a novel microcavitary hydrogel was proven to be effective for proliferation of chondrocytes and maintenance of chondrocytic phenotype. In present work, we further investigated whether the size of microcavity would affect the growth and the function of chondrocytes. By changing the stirring rate, gelatin microspheres in different sizes including small size (80–120 μm), middle size (150–200 μm) and large size (250–300 μm) were prepared. And then porcine chondrocytes were encapsulated into alginate hydrogel with various sizes of gelatin microspheres. Cell Counting Kit-8 (CCK-8), Live/dead staining and real-time PCR were used to analyze the effect of the pore size on cell proliferation and expression of specific chondrocytic genes. According to all the data, cells cultivated in microcavitary hydrogel, especially in small size, had preferable abilities of proliferation and higher expression of cartilaginous markers including type II collagen, aggrecan and cartilage oligomeric matrix protein (COMP). Furthermore, it was shown by western blot assay that the culture of chondrocytes in microcavitary hydrogel could improve the proliferation of cells potentially by inducing the Erk1/2-MAPK pathway. Taken together, this study demonstrated that chondrocytes favored microcavitary alginate hydrogel with pore size within the range of 80–120 μm for better growth and ECM synthesis, in which Erk1/2 pathway was involved. This culture system would be promising for cartilage tissue engineering. - Highlights: • A novel model with microcavitary structure was set up to study the interaction between cells and materials. • Microcavitary alginate hydrogel could enhance the proliferation of chondrocytes and promote the expression of cartilaginous genes as compared with plain alginate hydrogel. • Cells in microcavitary alginate hydrogel with pore size within the range of 80–120 μm were capable of better growth and ECM synthesis

  18. Subchondral Bone Plate Thickening Precedes Chondrocyte Apoptosis and Cartilage Degradation in Spontaneous Animal Models of Osteoarthritis

    OpenAIRE

    Zamli, Zaitunnatakhin; Robson Brown, Kate; Tarlton, John F.; Adams, Mike A.; Torlot, Georgina E.; Cartwright, Charlie; Cook, William A.; Vassilevskaja, Kristiina; Sharif, Mohammed

    2014-01-01

    Osteoarthritis (OA) is the most common joint disorder characterised by bone remodelling and cartilage degradation and associated with chondrocyte apoptosis. These processes were investigated at 10, 16, 24, and 30 weeks in Dunkin Hartley (DH) and Bristol Strain 2 (BS2) guinea pigs that develop OA spontaneously. Both strains had a more pronounced chondrocyte apoptosis, cartilage degradation, and subchondral bone changes in the medial than the lateral side of the tibia, and between strains, the ...

  19. Secondary chondrocyte-derived Ihh stimulates proliferation of periosteal cells during chick development.

    Science.gov (United States)

    Buxton, Paul G; Hall, Brian; Archer, Charles W; Francis-West, Philippa

    2003-10-01

    The development of the skull is characterised by its dependence upon epigenetic influences. One of the most important of these is secondary chondrogenesis, which occurs following ossification within certain membrane bone periostea, as a result of biomechanical articulation. We have studied the genesis, character and function of the secondary chondrocytes of the quadratojugal of the chick between embryonic days 11 and 14. Analysis of gene expression revealed that secondary chondrocytes formed coincident with Sox9 upregulation from a precursor population expressing Cbfa1/Runx2: a reversal of the normal sequence. Such secondary chondrocytes rapidly acquired a phenotype that is a compound of prehypertrophic and hypertrophic chondrocytes, exited from the cell cycle and upregulated Ihh. Pulse and pulse/chase experiments with BrdU confirmed the germinal region as the highly proliferative source of the secondary chondrocytes, which formed by division of chondrocyte-committed precursors. By blocking Hh signalling in explant cultures we show that the enhanced proliferation of the germinal region surrounding the secondary chondrocytes derives from this Ihh source. Additionally, in vitro studies on membrane bone periosteal cells (non-germinal region) demonstrated that these cells can also respond to Ihh, and do so both by enhanced proliferation and precocious osteogenesis. Despite the pro-osteogenic effects of Ihh on periosteal cell differentiation, mechanical articulation of the quadratojugal/quadrate joint in explant culture revealed a negative role for articulation in the regulation of osteocalcin by germinal region descendants. Thus, the mechanical stimulus that is the spur to secondary chondrocyte formation appears able to override the osteogenic influence of Ihh on the periosteum, but does not interfere with the cell cycle-promoting component of Hh signalling.

  20. Changes in the expression of collagen genes show two stages in chondrocyte differentiation in vitro

    OpenAIRE

    1988-01-01

    This report deals with the quantitation of both mRNA and transcription activity of type I collagen gene and of three cartilage-specific collagens (types II, IX, and X) during in vitro differentiation of chick chondrocytes. Differentiation was obtained by transferal to suspension culture of dedifferentiated cells passaged for 3 wk as adherent cells. The type I collagen mRNA, highly represented in the dedifferentiated cells, rapidly decreased during chondrocyte differentiation. On the contrary,...

  1. Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA

    Directory of Open Access Journals (Sweden)

    Jakob Naranda

    2017-03-01

    Full Text Available Background Cartilage tissue engineering is a fast-evolving field of biomedical engineering, in which the chondrocytes represent the most commonly used cell type. Since research in tissue engineering always consumes a lot of cells, simple and cheap isolation methods could form a powerful basis to boost such studies and enable their faster progress to the clinics. Isolated chondrocytes can be used for autologous chondrocyte implantation in cartilage repair, and are the base for valuable models to investigate cartilage phenotype preservation, as well as enable studies of molecular features, nature and scales of cellular responses to alterations in the cartilage tissue. Methods Isolation and consequent cultivation of primary human adult articular chondrocytes from the surgical waste obtained during total knee arthroplasty (TKA was performed. To evaluate the chondrogenic potential of the isolated cells, gene expression of collagen type 2 (COL2, collagen 1 (COL1 and aggrecan (ACAN was evaluated. Immunocytochemical staining of all mentioned proteins was performed to evaluate chondrocyte specific production. Results Cartilage specific gene expression of COL2 and ACAN has been shown that the proposed protocol leads to isolation of cells with a high chondrogenic potential, possibly even specific phenotype preservation up to the second passage. COL1 expression has confirmed the tendency of the isolated cells dedifferentiation into a fibroblast-like phenotype already in the second passage, which confirms previous findings that higher passages should be used with care in cartilage tissue engineering. To evaluate the effectiveness of our approach, immunocytochemical staining of the evaluated chondrocyte specific products was performed as well. Discussion In this study, we developed a protocol for isolation and consequent cultivation of primary human adult articular chondrocytes with the desired phenotype from the surgical waste obtained during TKA. TKA is a

  2. Dynamic compression of chondrocyte-agarose constructs reveals new candidate mechanosensitive genes.

    Directory of Open Access Journals (Sweden)

    Carole Bougault

    Full Text Available Articular cartilage is physiologically exposed to repeated loads. The mechanical properties of cartilage are due to its extracellular matrix, and homeostasis is maintained by the sole cell type found in cartilage, the chondrocyte. Although mechanical forces clearly control the functions of articular chondrocytes, the biochemical pathways that mediate cellular responses to mechanical stress have not been fully characterised. The aim of our study was to examine early molecular events triggered by dynamic compression in chondrocytes. We used an experimental system consisting of primary mouse chondrocytes embedded within an agarose hydrogel; embedded cells were pre-cultured for one week and subjected to short-term compression experiments. Using Western blots, we demonstrated that chondrocytes maintain a differentiated phenotype in this model system and reproduce typical chondrocyte-cartilage matrix interactions. We investigated the impact of dynamic compression on the phosphorylation state of signalling molecules and genome-wide gene expression. After 15 min of dynamic compression, we observed transient activation of ERK1/2 and p38 (members of the mitogen-activated protein kinase (MAPK pathways and Smad2/3 (members of the canonical transforming growth factor (TGF-β pathways. A microarray analysis performed on chondrocytes compressed for 30 min revealed that only 20 transcripts were modulated more than 2-fold. A less conservative list of 325 modulated genes included genes related to the MAPK and TGF-β pathways and/or known to be mechanosensitive in other biological contexts. Of these candidate mechanosensitive genes, 85% were down-regulated. Down-regulation may therefore represent a general control mechanism for a rapid response to dynamic compression. Furthermore, modulation of transcripts corresponding to different aspects of cellular physiology was observed, such as non-coding RNAs or primary cilium. This study provides new insight into how

  3. Role of Insulin-Transferrin-Selenium in Auricular Chondrocyte Proliferation and Engineered Cartilage Formation in Vitro

    Directory of Open Access Journals (Sweden)

    Xia Liu

    2014-01-01

    Full Text Available The goal of this study is to determine the effects of Insulin-Transferrin-Selenium (ITS on proliferation of auricular chondrocytes and formation of engineered cartilage in vitro. Pig auricular monolayer chondrocytes and chondrocyte pellets were cultured in media containing 1% ITS at different concentrations of fetal bovine serum (FBS, 10%, 6%, 2%, 0%, or 10% FBS alone as a control for four weeks. Parameters including cell proliferation in monolayer, wet weight, collagen type I/II/X (Col I, II, X and glycosaminoglycan (GAG expression, GAG content of pellets and gene expression associated with cartilage formation/dedifferentiation (lost cartilage phenotype/hypertrophy within the chondrocyte pellets were assessed. The results showed that chondrocytes proliferation rates increased when FBS concentrations increased (2%, 6%, 10% FBS in ITS supplemented groups. In addition, 1% ITS plus 10% FBS significantly promoted cell proliferation than 10% FBS alone. No chondrocytes grew in ITS alone medium. 1% ITS plus 10% FBS enhanced cartilage formation in terms of size, wet weight, cartilage specific matrices, and homogeneity, compared to 10% FBS alone group. Furthermore, ITS prevented engineered cartilage from dedifferentiation (i.e., higher index of Col II/Col I mRNA expression and expression of aggrecan and hypertrophy (i.e., lower mRNA expression of Col X and MMP13. In conclusion, our results indicated that ITS efficiently enhanced auricular chondrocytes proliferation, retained chondrogenic phenotypes, and promoted engineered cartilage formation when combined with FBS, which is potentially used as key supplementation in auricular chondrocytes and engineered cartilage culture.

  4. High throughput generated micro-aggregates of chondrocytes stimulate cartilage formation in vitro and in vivo

    Directory of Open Access Journals (Sweden)

    LS Moreira Teixeira

    2012-06-01

    Full Text Available Cell-based cartilage repair strategies such as matrix-induced autologous chondrocyte implantation (MACI could be improved by enhancing cell performance. We hypothesised that micro-aggregates of chondrocytes generated in high-throughput prior to implantation in a defect could stimulate cartilaginous matrix deposition and remodelling. To address this issue, we designed a micro-mould to enable controlled high-throughput formation of micro-aggregates. Morphology, stability, gene expression profiles and chondrogenic potential of micro-aggregates of human and bovine chondrocytes were evaluated and compared to single-cells cultured in micro-wells and in 3D after encapsulation in Dextran-Tyramine (Dex-TA hydrogels in vitro and in vivo. We successfully formed micro-aggregates of human and bovine chondrocytes with highly controlled size, stability and viability within 24 hours. Micro-aggregates of 100 cells presented a superior balance in Collagen type I and Collagen type II gene expression over single cells and micro-aggregates of 50 and 200 cells. Matrix metalloproteinases 1, 9 and 13 mRNA levels were decreased in micro-aggregates compared to single-cells. Histological and biochemical analysis demonstrated enhanced matrix deposition in constructs seeded with micro-aggregates cultured in vitro and in vivo, compared to single-cell seeded constructs. Whole genome microarray analysis and single gene expression profiles using human chondrocytes confirmed increased expression of cartilage-related genes when chondrocytes were cultured in micro-aggregates. In conclusion, we succeeded in controlled high-throughput formation of micro-aggregates of chondrocytes. Compared to single cell-seeded constructs, seeding of constructs with micro-aggregates greatly improved neo-cartilage formation. Therefore, micro-aggregation prior to chondrocyte implantation in current MACI procedures, may effectively accelerate hyaline cartilage formation.

  5. High throughput generated micro-aggregates of chondrocytes stimulate cartilage formation in vitro and in vivo.

    Science.gov (United States)

    Moreira Teixeira, L S; Leijten, J C H; Sobral, J; Jin, R; van Apeldoorn, A A; Feijen, J; van Blitterswijk, C; Dijkstra, P J; Karperien, M

    2012-06-05

    Cell-based cartilage repair strategies such as matrix-induced autologous chondrocyte implantation (MACI) could be improved by enhancing cell performance. We hypothesised that micro-aggregates of chondrocytes generated in high-throughput prior to implantation in a defect could stimulate cartilaginous matrix deposition and remodelling. To address this issue, we designed a micro-mould to enable controlled high-throughput formation of micro-aggregates. Morphology, stability, gene expression profiles and chondrogenic potential of micro-aggregates of human and bovine chondrocytes were evaluated and compared to single-cells cultured in micro-wells and in 3D after encapsulation in Dextran-Tyramine (Dex-TA) hydrogels in vitro and in vivo. We successfully formed micro-aggregates of human and bovine chondrocytes with highly controlled size, stability and viability within 24 hours. Micro-aggregates of 100 cells presented a superior balance in Collagen type I and Collagen type II gene expression over single cells and micro-aggregates of 50 and 200 cells. Matrix metalloproteinases 1, 9 and 13 mRNA levels were decreased in micro-aggregates compared to single-cells. Histological and biochemical analysis demonstrated enhanced matrix deposition in constructs seeded with micro-aggregates cultured in vitro and in vivo, compared to single-cell seeded constructs. Whole genome microarray analysis and single gene expression profiles using human chondrocytes confirmed increased expression of cartilage-related genes when chondrocytes were cultured in micro-aggregates. In conclusion, we succeeded in controlled high-throughput formation of micro-aggregates of chondrocytes. Compared to single cell-seeded constructs, seeding of constructs with micro-aggregates greatly improved neo-cartilage formation. Therefore, micro-aggregation prior to chondrocyte implantation in current MACI procedures, may effectively accelerate hyaline cartilage formation.

  6. Effect of microcavitary alginate hydrogel with different pore sizes on chondrocyte culture for cartilage tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Zeng, Lei; Yao, Yongchang [School of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 (China); National Engineering Research Center for Tissue Restoration and Reconstruction, Guangzhou 510006 (China); Wang, Dong-an, E-mail: DAWang@ntu.edu.sg [National Engineering Research Center for Tissue Restoration and Reconstruction, Guangzhou 510006 (China); Division of Bioengineering, School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637457 (Singapore); Chen, Xiaofeng, E-mail: chenxf@scut.edu.cn [School of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 (China); National Engineering Research Center for Tissue Restoration and Reconstruction, Guangzhou 510006 (China)

    2014-01-01

    In our previous work, a novel microcavitary hydrogel was proven to be effective for proliferation of chondrocytes and maintenance of chondrocytic phenotype. In present work, we further investigated whether the size of microcavity would affect the growth and the function of chondrocytes. By changing the stirring rate, gelatin microspheres in different sizes including small size (80–120 μm), middle size (150–200 μm) and large size (250–300 μm) were prepared. And then porcine chondrocytes were encapsulated into alginate hydrogel with various sizes of gelatin microspheres. Cell Counting Kit-8 (CCK-8), Live/dead staining and real-time PCR were used to analyze the effect of the pore size on cell proliferation and expression of specific chondrocytic genes. According to all the data, cells cultivated in microcavitary hydrogel, especially in small size, had preferable abilities of proliferation and higher expression of cartilaginous markers including type II collagen, aggrecan and cartilage oligomeric matrix protein (COMP). Furthermore, it was shown by western blot assay that the culture of chondrocytes in microcavitary hydrogel could improve the proliferation of cells potentially by inducing the Erk1/2-MAPK pathway. Taken together, this study demonstrated that chondrocytes favored microcavitary alginate hydrogel with pore size within the range of 80–120 μm for better growth and ECM synthesis, in which Erk1/2 pathway was involved. This culture system would be promising for cartilage tissue engineering. - Highlights: • A novel model with microcavitary structure was set up to study the interaction between cells and materials. • Microcavitary alginate hydrogel could enhance the proliferation of chondrocytes and promote the expression of cartilaginous genes as compared with plain alginate hydrogel. • Cells in microcavitary alginate hydrogel with pore size within the range of 80–120 μm were capable of better growth and ECM synthesis.

  7. Haemodynamics in axillobifemoral bypass grafts

    NARCIS (Netherlands)

    C.H. Wittens

    1992-01-01

    textabstractThis thesis is based on four publications on the subject of graft configuration and haemodynamics in axillobifemoral bypass grafts: 1. A clinical evaluation of 17 patients with axillobifemoral bypass graft operations, performed for various indications. Two important observations were

  8. NF-kappaB specifically activates BMP-2 gene expression in growth plate chondrocytes in vivo and in a chondrocyte cell line in vitro.

    Science.gov (United States)

    Feng, Jian Q; Xing, Lianping; Zhang, Jiang-Hong; Zhao, Ming; Horn, Diane; Chan, Jeannie; Boyce, Brendan F; Harris, Stephen E; Mundy, Gregory R; Chen, Di

    2003-08-01

    Bone morphogenetic protein-2 (BMP-2) regulates growth plate chondrogenesis during development and postnatal bone growth, but the control mechanisms of BMP-2 expression in growth plate chondrocytes are unknown. Here we have used both in vitro and in vivo approaches to demonstrate that transcription factor, NF-kappaB, regulates BMP-2 gene expression in chondrocytes. Two putative NF-kappaB response elements were found in the -2712/+165 region of the BMP-2 gene. Cotransfection of mutant I-kappaBalpha expression plasmids with BMP-2 promoter-luciferase reporters into TMC-23 chondrocyte cell line suppressed BMP-2 transcription. Mutations in NF-kappaB response elements in the BMP-2 gene lead to decreases in BMP-2 promoter activity. Electrophoretic mobility shift assay using nuclear extracts from TMC-23 chondrocytic cells revealed that the NF-kappaB subunits p50 and p65 bound to the NF-kappaB response elements of the BMP-2 gene. Thus, NF-kappaB may positively regulate BMP-2 gene transcription. Consistent with these findings, expression of BMP-2 mRNA was significantly reduced in growth plate chondrocytes in NF-kappaB p50/p52 dKO mice, which associated with decreased numbers of 5-bromo-2'-deoxyuridine (BrdUrd)-positive cells in the proliferating zone of growth plate in these mice. Therefore, in postnatal growth plate chondrocytes, expression of BMP-2 is regulated by NF-kappaB, which may play an important role in chondrogenesis.

  9. Chondrocytes and stem cells in 3D-bioprinted structures create human cartilage in vivo.

    Science.gov (United States)

    Apelgren, Peter; Amoroso, Matteo; Lindahl, Anders; Brantsing, Camilla; Rotter, Nicole; Gatenholm, Paul; Kölby, Lars

    2017-01-01

    Cartilage repair and replacement is a major challenge in plastic reconstructive surgery. The development of a process capable of creating a patient-specific cartilage framework would be a major breakthrough. Here, we described methods for creating human cartilage in vivo and quantitatively assessing the proliferative capacity and cartilage-formation ability in mono- and co-cultures of human chondrocytes and human mesenchymal stem cells in a three-dimensional (3D)-bioprinted hydrogel scaffold. The 3D-bioprinted constructs (5 × 5 × 1.2 mm) were produced using nanofibrillated cellulose and alginate in combination with human chondrocytes and human mesenchymal stem cells using a 3D-extrusion bioprinter. Immediately following bioprinting, the constructs were implanted subcutaneously on the back of 48 nude mice and explanted after 30 and 60 days, respectively, for morphological and immunohistochemical examination. During explantation, the constructs were easy to handle, and the majority had retained their macroscopic grid appearance. Constructs consisting of human nasal chondrocytes showed good proliferation ability, with 17.2% of the surface areas covered with proliferating chondrocytes after 60 days. In constructs comprising a mixture of chondrocytes and stem cells, an additional proliferative effect was observed involving chondrocyte production of glycosaminoglycans and type 2 collagen. This clinically highly relevant study revealed 3D bioprinting as a promising technology for the creation of human cartilage.

  10. Chondrocytes and stem cells in 3D-bioprinted structures create human cartilage in vivo.

    Directory of Open Access Journals (Sweden)

    Peter Apelgren

    Full Text Available Cartilage repair and replacement is a major challenge in plastic reconstructive surgery. The development of a process capable of creating a patient-specific cartilage framework would be a major breakthrough. Here, we described methods for creating human cartilage in vivo and quantitatively assessing the proliferative capacity and cartilage-formation ability in mono- and co-cultures of human chondrocytes and human mesenchymal stem cells in a three-dimensional (3D-bioprinted hydrogel scaffold. The 3D-bioprinted constructs (5 × 5 × 1.2 mm were produced using nanofibrillated cellulose and alginate in combination with human chondrocytes and human mesenchymal stem cells using a 3D-extrusion bioprinter. Immediately following bioprinting, the constructs were implanted subcutaneously on the back of 48 nude mice and explanted after 30 and 60 days, respectively, for morphological and immunohistochemical examination. During explantation, the constructs were easy to handle, and the majority had retained their macroscopic grid appearance. Constructs consisting of human nasal chondrocytes showed good proliferation ability, with 17.2% of the surface areas covered with proliferating chondrocytes after 60 days. In constructs comprising a mixture of chondrocytes and stem cells, an additional proliferative effect was observed involving chondrocyte production of glycosaminoglycans and type 2 collagen. This clinically highly relevant study revealed 3D bioprinting as a promising technology for the creation of human cartilage.

  11. The Ihh signal is essential for regulating proliferation and hypertrophy of cultured chicken chondrocytes.

    Science.gov (United States)

    Ma, R S; Zhou, Z L; Luo, J W; Zhang, H; Hou, J F

    2013-10-01

    The Indian hedgehog (Ihh) signal plays a vital role in regulating proliferation and hypertrophy of chondrocytes. To investigate its function in postnatal chicken (Gallus gallus) chondrocytes, cyclopamine was used to inhibit Ihh signaling. The MTT and ALP assays revealed the downgrade-proliferation and upgrade-differentiation of chondrocytes. To further elucidate the mechanism, the mRNA expression levels of Ihh, parathyroid hormone related protein (PTHrP), Gli-2, Bcl-2, Bone Morphogenetic Protein 6 (BMP-6), type X collagen (Col X) and type II collagen (Col II) were detected by quantitative real-time RT-PCR analysis, and the protein expressions of Ihh, Col X, and Col II were determined using Western blot analysis. After the Ihh signal was blocked, chondrocytes demonstrated high expression levels of PTHrP and Col X and low levels of Gli-2, BMP-6, Bcl-2 and Col II although Ihh expression was increased. Based on these results, the Ihh signal is essential for balancing chicken chondrocyte proliferation and hypertrophy, and the regulatory function of PTHrP acts in an Ihh-dependent manner. Furthermore, BMP-6 and Bcl-2 played roles in maintaining the development of chondrocytes and may be downstream regulatory factors of Ihh signaling. © 2013.

  12. Enhanced chondrocyte culture and growth on biologically inspired nanofibrous cell culture dishes.

    Science.gov (United States)

    Bhardwaj, Garima; Webster, Thomas J

    2016-01-01

    Chondral and osteochondral defects affect a large number of people in which treatment options are currently limited. Due to its ability to mimic the natural nanofibrous structure of cartilage, this current in vitro study aimed at introducing a new scaffold, called XanoMatrix™, for cartilage regeneration. In addition, this same scaffold is introduced here as a new substrate onto which to study chondrocyte functions. Current studies on chondrocyte functions are limited due to nonbiologically inspired cell culture substrates. With its polyethylene terephthalate and cellulose acetate composition, good mechanical properties and nanofibrous structure resembling an extracellular matrix, XanoMatrix offers an ideal surface for chondrocyte growth and proliferation. This current study demonstrated that the XanoMatrix scaffolds promote chondrocyte growth and proliferation as compared with the Corning and Falcon surfaces normally used for chondrocyte cell culture. The XanoMatrix scaffolds also have greater hydrophobicity, three-dimensional surface area, and greater tensile strength, making them ideal candidates for alternative treatment options for chondral and osteochondral defects as well as cell culture substrates to study chondrocyte functions.

  13. Histone Deacetylase 3 Suppresses Erk Phosphorylation and Matrix Metalloproteinase (Mmp)-13 Activity in Chondrocytes

    Science.gov (United States)

    Carpio, Lomeli R.; Bradley, Elizabeth W.; Westendorf, Jennifer J.

    2017-01-01

    Histone deacetylase inhibitors are emerging therapies for many diseases including cancers and neurological disorders; however, these drugs are teratogens to the developing skeleton. Hdac3 is essential for proper endochondral ossification as its deletion in chondrocytes increases cytokine signaling and the expression of matrix remodeling enzymes. Here we explored the mechanism by which Hdac3 controls Mmp13 expression in chondrocytes. In Hdac3-depleted chondrocytes, Erk1/2 as well as its downstream substrate, Runx2, were hyperphosphorylated as a result of decreased expression and activity of the Erk1/2 specific phosphatase, Dusp6. Erk1/2 kinase inhibitors and Dusp6 adenoviruses reduced Mmp13 expression and partially rescued matrix production in Hdac3-deficient chondrocytes. Postnatal chondrocyte-specific deletion of Hdac3 with an inducible Col2a1-Cre caused premature production of pErk1/2 and Mmp13 in the growth plate. Thus, Hdac3 controls the temporal and spatial expression of tissue-remodeling genes in chondrocytes to ensure proper endochondral ossification during development. PMID:27662443

  14. Passaged adult chondrocytes can form engineered cartilage with functional mechanical properties: a canine model.

    Science.gov (United States)

    Ng, Kenneth W; Lima, Eric G; Bian, Liming; O'Conor, Christopher J; Jayabalan, Prakash S; Stoker, Aaron M; Kuroki, Keiichi; Cook, Cristi R; Ateshian, Gerard A; Cook, James L; Hung, Clark T

    2010-03-01

    It was hypothesized that previously optimized serum-free culture conditions for juvenile bovine chondrocytes could be adapted to generate engineered cartilage with physiologic mechanical properties in a preclinical, adult canine model. Primary or passaged (using growth factors) adult chondrocytes from three adult dogs were encapsulated in agarose, and cultured in serum-free media with transforming growth factor-beta3. After 28 days in culture, engineered cartilage formed by primary chondrocytes exhibited only small increases in glycosaminoglycan content. However, all passaged chondrocytes on day 28 elaborated a cartilage matrix with compressive properties and glycosaminoglycan content in the range of native adult canine cartilage values. A preliminary biocompatibility study utilizing chondral and osteochondral constructs showed no gross or histological signs of rejection, with all implanted constructs showing excellent integration with surrounding cartilage and subchondral bone. This study demonstrates that adult canine chondrocytes can form a mechanically functional, biocompatible engineered cartilage tissue under optimized culture conditions. The encouraging findings of this work highlight the potential for tissue engineering strategies using adult chondrocytes in the clinical treatment of cartilage defects.

  15. Efficiency of Human Epiphyseal Chondrocytes with Differential Replication Numbers for Cellular Therapy Products

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    Michiyo Nasu

    2016-01-01

    Full Text Available The cell-based therapy for cartilage or bone requires a large number of cells; serial passages of chondrocytes are, therefore, needed. However, fates of expanded chondrocytes from extra fingers remain unclarified. The chondrocytes from human epiphyses morphologically changed from small polygonal cells to bipolar elongated spindle cells and to large polygonal cells with degeneration at early passages. Gene of type II collagen was expressed in the cells only at a primary culture (Passage 0 and Passage 1 (P1 cells. The nodules by implantation of P0 to P8 cells were composed of cartilage and perichondrium. The cartilage consisted of chondrocytes with round nuclei and type II collagen-positive matrix, and the perichondrium consisted of spindle cells with type I collage-positive matrix. The cartilage and perichondrium developed to bone with marrow cavity through enchondral ossification. Chondrogenesis and osteogenesis by epiphyseal chondrocytes depended on replication number in culture. It is noteworthy to take population doubling level in correlation with pharmaceutical efficacy into consideration when we use chondrocytes for cell-based therapies.

  16. Functional regeneration of ligament-bone interface using a triphasic silk-based graft.

    Science.gov (United States)

    Li, Hongguo; Fan, Jiabing; Sun, Liguo; Liu, Xincheng; Cheng, Pengzhen; Fan, Hongbin

    2016-11-01

    The biodegradable silk-based scaffold with unique mechanical property and biocompatibility represents a favorable ligamentous graft for tissue-engineering anterior cruciate ligament (ACL) reconstruction. However, the low efficiency of ligament-bone interface restoration barriers the isotropic silk graft to common ACL therapeutics. To enhance the regeneration of the silk-mediated interface, we developed a specialized stratification approach implementing a sequential modification on isotropic silk to constitute a triphasic silk-based graft in which three regions respectively referring to ligament, cartilage and bone layers of interface were divided, followed by respective biomaterial coating. Furthermore, three types of cells including bone marrow mesenchymal stem cells (BMSCs), chondrocytes and osteoblasts were respectively seeded on the ligament, cartilage and bone region of the triphasic silk graft, and the cell/scaffold complex was rolled up as a multilayered graft mimicking the stratified structure of native ligament-bone interface. In vitro, the trilineage cells loaded on the triphasic silk scaffold revealed a high proliferative capacity as well as enhanced differentiation ability into their corresponding cell lineage. 24 weeks postoperatively after the construct was implanted to repair the ACL defect in rabbit model, the silk-based ligamentous graft exhibited the enhancement of osseointegration detected by a robust pullout force and formation of three-layered structure along with conspicuously corresponding matrix deposition via micro-CT and histological analysis. These findings potentially broaden the application of silk-based ligamentous graft for ACL reconstruction and further large animal study. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Primary renal graft thrombosis

    NARCIS (Netherlands)

    Bakir, N; Sluiter, WJ; Ploeg, RJ; van Son, WJ; Tegzess, Adam

    Background. Renal allograft thrombosis is a serious complication of kidney transplantation that ultimately leads to graft loss. Its association with acute and hyperacute rejection is well documented; however, in a large proportion of patients the precise cause remains obscure. The exact incidence

  18. Carnosol Inhibits Pro-Inflammatory and Catabolic Mediators of Cartilage Breakdown in Human Osteoarthritic Chondrocytes and Mediates Cross-Talk between Subchondral Bone Osteoblasts and Chondrocytes.

    Directory of Open Access Journals (Sweden)

    Christelle Sanchez

    Full Text Available The aim of this work was to evaluate the effects of carnosol, a rosemary polyphenol, on pro-inflammatory and catabolic mediators of cartilage breakdown in chondrocytes and via bone-cartilage crosstalk.Osteoarthritic (OA human chondrocytes were cultured in alginate beads for 4 days in presence or absence of carnosol (6 nM to 9 μM. The production of aggrecan, matrix metalloproteinase (MMP-3, tissue inhibitor of metalloproteinase (TIMP-1, interleukin (IL-6 and nitric oxide (NO and the expression of type II collagen and ADAMTS-4 and -5 were analyzed. Human osteoblasts from sclerotic (SC or non-sclerotic (NSC subchondral bone were cultured for 3 days in presence or absence of carnosol before co-culture with chondrocytes. Chondrocyte gene expression was analyzed after 4 days of co-culture.In chondrocytes, type II collagen expression was significantly enhanced in the presence of 3 μM carnosol (p = 0.008. MMP-3, IL-6, NO production and ADAMTS-4 expression were down-regulated in a concentration-dependent manner by carnosol (p<0.01. TIMP-1 production was slightly increased at 3 μM (p = 0.02 and ADAMTS-5 expression was decreased from 0.2 to 9 μM carnosol (p<0.05. IL-6 and PGE2 production was reduced in the presence of carnosol in both SC and NSC osteoblasts while alkaline phosphatase activity was not changed. In co-culture experiments preincubation of NSC and SC osteoblasts wih carnosol resulted in similar effects to incubation with anti-IL-6 antibody, namely a significant increase in aggrecan and decrease in MMP-3, ADAMTS-4 and -5 gene expression by chondrocytes.Carnosol showed potent inhibition of pro-inflammatory and catabolic mediators of cartilage breakdown in chondrocytes. Inhibition of matrix degradation and enhancement of formation was observed in chondrocytes cocultured with subchondral osteoblasts preincubated with carnosol indicating a cross-talk between these two cellular compartments, potentially mediated via inhibition of IL-6 in

  19. Aortic Graft Infection Secondary to Iatrogenic Transcolonic Graft Malposition.

    Science.gov (United States)

    Blank, Jacqueline J; Rothstein, Abby E; Lee, Cheong Jun; Malinowski, Michael J; Lewis, Brian D; Ridolfi, Timothy J; Otterson, Mary F

    2018-01-01

    Aortic graft infections are a rare but devastating complication of aortic revascularization. Often infections occur due to contamination at the time of surgery. Iatrogenic misplacement of the limbs of an aortobifemoral graft is exceedingly rare, and principles of evaluation and treatment are not well defined. We report 2 cases of aortobifemoral bypass graft malposition through the colon. Case 1 is a 54-year-old male who underwent aortobifemoral bypass grafting for acute limb ischemia. He had previously undergone a partial sigmoid colectomy for diverticulitis. Approximately 6 months after vascular surgery, he presented with an occult graft infection. Preoperative imaging and intraoperative findings were consistent with graft placement through the sigmoid colon. Case 2 is a 60-year-old male who underwent aortobifemoral bypass grafting due to a nonhealing wound after toe amputation. His postoperative course was complicated by pneumonia, bacteremia thought to be secondary to the pneumonia, general malaise, and persistent fevers. Approximately 10 weeks after the vascular surgery, he presented with imaging and intraoperative findings of graft malposition through the cecum. Aortic graft infection is usually caused by surgical contamination and presents as an indolent infection. Case 1 presented as such; Case 2 presented more acutely. Both grafts were iatrogenically misplaced through the colon at the index operation. The patients underwent extra-anatomic bypass and graft explantation and subsequently recovered.

  20. ACTIVITY OF CANONICAL WNT SIGNAL SYSTEM IN HYALINE CARTILAGE ARTICULAR CHONDROCYTES IN PROCESS OF SYNOVIAL JOINT DEVELOPMENT

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    A.O. Molotkov

    2009-03-01

    Full Text Available Canonical and non-canonical Wnt systems are essential regulators of chondrogenesis and bone development. However, the roles of these systems in synovial joint development are not well studied. To determine if canonical Wnt system is active in developing articular chondrocytes we used immunohistochemistry for в-galactosidase and doublecortin (cell-type specific marker for articular chondrocytes to double label sections through joint regions of E14.5, E18.5, P10 and adult mice. Here the following results are presented. Canonical Wnt signal system does not work in developing articular chondrocytes at early embryonic stages (E14.5; it is active in the articular chondrocytes at late embryonic stages (E16.5-E18.5 and during postnatal development (P7-P10, but is turned off again in the adult articular chondrocytes. These results suggest that canonical Wnt signaling is being regulated during articular chondrocytes differentiation and joint formation.

  1. Type II collagen peptide is able to accelerate embryonic chondrocyte differentiation: an association with articular cartilage matrix resorption in osteoarthrosis

    Directory of Open Access Journals (Sweden)

    Elena Vasil'evna Chetina

    2010-01-01

    Conclusion. The effect of CP on gene expression and collagen decomposition activity depends on the morphotype of embryonic chondrocytes. Lack of effect of CP on collagen decomposition activity in both the embryonic hypertrophic chondrocytes and the cartilage explants from OA patients supports the hypothesis that the hypertrophic morphotype is a dominant morphotype of articular chondrocytes in OA. Moreover, collagen decomposition products can be involved in the resorption of matrix in OA and in the maintenance of chronic nature of the pathology.

  2. Prognosis and continuity of child mental health problems from preschool to primary school: results of a four-year longitudinal study.

    Science.gov (United States)

    Beyer, Thomas; Postert, Christian; Müller, Jörg M; Furniss, Tilman

    2012-08-01

    In a four-year longitudinal study, changes in and continuity of behavioral and emotional problems were examined in 814 subjects from kindergarten to primary school. Mental health problems were assessed by means of the Child Behavior Checklist (CBCL). The distribution of the CBCL broadband groups revealed a high level of continuity of internalizing symptoms over the four-year period and a shift from externalizing symptoms at baseline towards a combination of internalizing and externalizing symptoms at follow-up. The presence of mental health problems at follow-up was correlated with gender (higher amongst boys), pre-existing mental health problems at baseline, and separation or divorce of the parents, but not with single-family status or the age and educational level of the mother. The increasing number of children with a combination of internalizing and externalizing symptoms demonstrates the increasing complexity of child mental health problems in the developmental span from preschool age to school age.

  3. The effect of nurse education on the self-esteem and assertiveness of nursing students: A four-year longitudinal study.

    Science.gov (United States)

    İlhan, Nesrin; Sukut, Özge; Akhan, Latife Utaş; Batmaz, Makbule

    2016-04-01

    The nurse education program affects many aspects of personal development. Nursing students gain skills in critical thinking and analysis and also develop communication and management skills. The four-year program may also have an effect on students' assertiveness and self-esteem. This study was conducted to determine the impact of the four-year higher nursing school educational program on students' self-esteem and assertiveness. Descriptive longitudinal design. The study took place at a Foundation University in Istanbul, Turkey over the period 2006-2010. The students' levels of self-esteem and assertiveness were assessed at the beginning and end of the first, second, third and fourth years of the program. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. The Socio-Cultural and Leadership Experiences of Latina Four-Year College and University Presidents: A Traves de sus Voces (Through Their Voices)

    Science.gov (United States)

    Maes, Johanna B.

    2012-01-01

    The Latina population in the United States is one of the largest of all racial and ethnic groups, and it is expected to grow exponentially within the next forty years. Despite these large numbers of Latinas in the U.S., there is a disparity with this population who are leading our nation's four-year colleges and universities. A reason for…

  5. Normal age-related viscoelastic properties of chondrons and chondrocytes isolated from rabbit knee

    Institute of Scientific and Technical Information of China (English)

    DUAN Wang-ping; SUN Zhen-wei; LI Qi; LI Chun-jiang; WANG Li; CHEN Wei-yi; Jennifer Tickner; ZHENG Ming-hao; WEI Xiao-chun

    2012-01-01

    Background The mechanical microenvironment of the chondrocytes plays an important role in cartilage homeostasis and in the health of the joint.The pericellular matrix,cellular membrane of the chondrocytes,and their cytoskeletal structures are key elements in the mechanical environment.The aims of this study are to measure the viscoelastic properties of isolated chondrons and chondrocytes from rabbit knee cartilage using micropipette aspiration and to determine the effect of aging on these properties.Methods Three age groups of rabbit knees were evaluated:(1) young (2 months,n=10);(2) adult (8 months,n=10);and (3) old (31 months,n=10).Chondrocytes were isolated from the right knee cartilage and chondrons were isolated from left knees using enzymatic methods.Micropipette aspiration combined with a standard linear viscoelastic solid model was used to quantify changes in the viscoelastic properties of chondrons and chondrocytes within 2 hours of isolation.The morphology and structure of isolated chondrons were evaluated by optical microscope using hematoxylin and eosin staining and collagen-6 immunofluorescence staining.Results In response to an applied constant 0.3-0.4 kPa of negative pressure,all chondrocytes exhibited standard linear viscoelastic solid properties.Model predictions of the creep data showed that the average equilibrium modulus (E∞),instantaneous modulus (E0).and apparent viscosity (μ) of old chondrocytes was significantly lower than the young and adult chondrocytes (P<0.001);however,no difference was found between young and adult chondrocytes (P>0.05).The adult and old chondrons generally possessed a thicker pericellular matrix (PCM) with more enclosed cells.The young and adult chondrons exhibited the same viscoelastic creep behavior under a greater applied pressure (1.0-1.1kPa) without the deformation seen in the old chondrons.The viscoelastic properties (E∞,E0,and u) of young and adult chondrons were significantly greater than that observed

  6. A four-year follow-up controlled study of stress response and symptom persistence in Brazilian children and adolescents with attention deficit disorder and hyperactivity (ADHD).

    Science.gov (United States)

    Palma, Sonia Maria Motta; Natale, Ana Carolina Motta Palma; Calil, Helena Maria

    2015-12-15

    This study evaluated children and adolescents with Attention Deficit Disorder andHyperactivity Disorder (ADHD), reassessing them at a four-year follow-up. Their cortisol response to a stress stimulus was measured twice. ADHD symptom persistence, development of comorbidities, and psychostimulant usage were also reassessed. The initial sample consisted of 38 ADHD patients and 38 healthy controls, age ranging 6-14. At the follow-up, there were 37 ADHD patients and 22 healthy controls, age ranging 10-18. ADHD was classified as persistent if the patients fulfilled all DSM IV criteria for syndromic or subthreshold or had functional impairment. Salivary cortisol samples were collected prior to the application of a cognitive stressor (Continuous Performance Test - CPT), and at three time intervals afterwards at baseline and at the follow-up. Their reassessment showed that 75% had persistent symptoms, psychiatric comorbidities (oppositional defiant and behavioral disorders), functional and academic impairement. Only seven patients were on medication. The ADHD group's cortisol levels were lower than those measured four years earlier, but cortisol concentrations were similar for both ADHD and control groups at the four-year follow-up. The cortisol results suggest that HPA axis reactivity could be a marker differentiating ADHD from ADHD with comorbidities. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  7. Incidence rate of falls and its risk factors in patients with rheumatoid arthritis compared to controls: Four years of the TOMORROW study.

    Science.gov (United States)

    Mamoto, Kenji; Inui, Kentaro; Okano, Tadashi; Sugioka, Yuko; Tada, Masahiro; Koike, Tatsuya; Nakamura, Hiroaki

    2017-01-01

    Patients with rheumatoid arthritis (RA) have been recognized to experience falls frequently due to functional disabilities. The aim of this study was to prospectively investigate factors influencing falls in patients with RA compared to controls. We compared the frequency of falls in 208 RA patients and 205 age- and sex-matched volunteers for four years and analyzed risk factors for falls in RA patients using multivariate regression analysis. No significant difference in the incidence rate of falls (/person-year) between patients with RA (median [interquartile range]: 0 [0, 0.5]) and controls (0 [0, 0.5]) was evident during four years. Logistic regression analysis identified age, sex, body mass index, history of falls, and lower limb implant at baseline as significant risk factors for falls. The highest quartile of anti-CCP antibody level (>300.6 U/ml) was the strongest predictor for multiple falls (odds ratio, 2.97; 95% confidence interval, 1.12-7.91, p = 0.029) among RA patients. During four years we could not observe the higher incidence rate of falls in RA patients compared to controls in our cohort. Subjects with a higher titer of anti-CCP antibody might be at higher risk of frequent falls among RA patients.

  8. Bullying and victimisation are common in four-year-old children and are associated with somatic symptoms and conduct and peer problems.

    Science.gov (United States)

    Ilola, Anna-Marja; Lempinen, Lotta; Huttunen, Jukka; Ristkari, Terja; Sourander, Andre

    2016-05-01

    There are few population-based studies on bullying behaviour among preschool children. The aims of the study were to investigate the prevalence of bullying behaviour among four-year-old children, as reported by their parents, the prevalence of types of bullying behaviour and the associations between bullying behaviour and psychosocial factors. This study was based on a population-based study sample of 931 children who attended their check-up at a child health clinic at four years of age. Parents completed the questionnaire about their child's bullying behaviour and risk factors during the check-up. Bullying behaviour, especially being both a bully and a victim, was a common phenomenon among four-year-old children. Being a bully or both a bully and victim were most strongly associated with conduct problems, while being a victim was associated with somatic symptoms and peer problems. Bullying behaviour was frequently found in preschool children and associated with a wide range of other problems, which indicate that routine checking of bullying behaviour should be included in child health clinic check-ups. Bullying prevention programmes are usually targeted at school-aged children, but this study highlights the importance of focusing already on preschool children. ©2016 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.

  9. The long-term association of OCD and depression and its moderators: A four-year follow up study in a large clinical sample.

    Science.gov (United States)

    Tibi, L; van Oppen, P; van Balkom, A J L M; Eikelenboom, M; Rickelt, J; Schruers, K R J; Anholt, G E

    2017-07-01

    Depression is the most common comorbidity in obsessive-compulsive disorder (OCD). However, the mechanisms of depressive comorbidity in OCD are poorly understood. We assessed the directionality and moderators of the OCD-depression association over time in a large, prospective clinical sample of OCD patients. Data were drawn from 382 OCD patients participating at the Netherlands Obsessive-Compulsive Disorder Association (NOCDA) study. Cross-lagged, structural equation modeling analyses were used to assess the temporal association between OCD and depressive symptoms. Assessments were conducted at baseline, two-year and four-year follow up. Cognitive and interpersonal moderators of the prospective association between OCD and depressive symptoms were tested. Cross-lagged analyses demonstrated that OCD predicts depressive symptoms at two-year follow up and not vice a versa. This relationship disappeared at four-year follow up. Secure attachment style moderated the prospective association between OCD and depression. Depressive comorbidity in OCD might constitute a functional consequence of the incapacitating OCD symptoms. Both OCD and depression symptoms demonstrated strong stability effects between two-year and four-year follow up, which may explain the lack of association between them in that period. Among OCD patients, secure attachment represents a buffer against future depressive symptoms. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  10. Biological and Chemical Removal of Primary Cilia Affects Mechanical Activation of Chondrogenesis Markers in Chondroprogenitors and Hypertrophic Chondrocytes.

    Science.gov (United States)

    Deren, Matthew E; Yang, Xu; Guan, Yingjie; Chen, Qian

    2016-02-04

    Chondroprogenitors and hypertrophic chondrocytes, which are the first and last stages of the chondrocyte differentiation process, respectively, are sensitive to mechanical signals. We hypothesize that the mechanical sensitivity of these cells depends on the cell surface primary cilia. To test this hypothesis, we removed the primary cilia by biological means with transfection with intraflagellar transport protein 88 (IFT88) siRNA or by chemical means with chloral hydrate treatment. Transfection of IFT88 siRNA significantly reduced the percentage of ciliated cells in both chondroprogenitor ATDC5 cells as well as primary hypertrophic chondrocytes. Cyclic loading (1 Hz, 10% matrix deformation) of ATDC5 cells in three-dimensional (3D) culture stimulates the mRNA levels of chondrogenesis marker Type II collagen (Col II), hypertrophic chondrocyte marker Type X collagen (Col X), and a molecular regulator of chondrogenesis and chondrocyte hypertrophy bone morphogenetic protein 2 (BMP-2). The reduction of ciliated chondroprogenitors abolishes mechanical stimulation of Col II, Col X, and BMP-2. In contrast, cyclic loading stimulates Col X mRNA levels in hypertrophic chondrocytes, but not those of Col II and BMP-2. Both biological and chemical reduction of ciliated hypertrophic chondrocytes reduced but failed to abolish mechanical stimulation of Col X mRNA levels. Thus, primary cilia play a major role in mechanical stimulation of chondrogenesis and chondrocyte hypertrophy in chondroprogenitor cells and at least a partial role in hypertrophic chondrocytes.

  11. Biological and Chemical Removal of Primary Cilia Affects Mechanical Activation of Chondrogenesis Markers in Chondroprogenitors and Hypertrophic Chondrocytes

    Directory of Open Access Journals (Sweden)

    Matthew E. Deren

    2016-02-01

    Full Text Available Chondroprogenitors and hypertrophic chondrocytes, which are the first and last stages of the chondrocyte differentiation process, respectively, are sensitive to mechanical signals. We hypothesize that the mechanical sensitivity of these cells depends on the cell surface primary cilia. To test this hypothesis, we removed the primary cilia by biological means with transfection with intraflagellar transport protein 88 (IFT88 siRNA or by chemical means with chloral hydrate treatment. Transfection of IFT88 siRNA significantly reduced the percentage of ciliated cells in both chondroprogenitor ATDC5 cells as well as primary hypertrophic chondrocytes. Cyclic loading (1 Hz, 10% matrix deformation of ATDC5 cells in three-dimensional (3D culture stimulates the mRNA levels of chondrogenesis marker Type II collagen (Col II, hypertrophic chondrocyte marker Type X collagen (Col X, and a molecular regulator of chondrogenesis and chondrocyte hypertrophy bone morphogenetic protein 2 (BMP-2. The reduction of ciliated chondroprogenitors abolishes mechanical stimulation of Col II, Col X, and BMP-2. In contrast, cyclic loading stimulates Col X mRNA levels in hypertrophic chondrocytes, but not those of Col II and BMP-2. Both biological and chemical reduction of ciliated hypertrophic chondrocytes reduced but failed to abolish mechanical stimulation of Col X mRNA levels. Thus, primary cilia play a major role in mechanical stimulation of chondrogenesis and chondrocyte hypertrophy in chondroprogenitor cells and at least a partial role in hypertrophic chondrocytes.

  12. The effects of monosodium urate monohydrate crystals on chondrocyte viability and function: implications for development of cartilage damage in gout.

    Science.gov (United States)

    Chhana, Ashika; Callon, Karen E; Pool, Bregina; Naot, Dorit; Gamble, Gregory D; Dray, Michael; Pitto, Rocco; Bentley, Jarome; McQueen, Fiona M; Cornish, Jillian; Dalbeth, Nicola

    2013-12-01

    Cartilage damage is frequently observed in advanced destructive gout. The aim of our study was to investigate the effects of monosodium urate monohydrate (MSU) crystals on chondrocyte viability and function. The alamarBlue assay and flow cytometry were used to assess the viability of primary human chondrocytes and cartilage explants following culture with MSU crystals. The number of dead chondrocytes in cartilage explants cultured with MSU crystals was quantified. Real-time PCR was used to determine changes in the relative mRNA expression levels of chondrocytic genes. The histological appearance of cartilage in joints affected by gout was also examined. MSU crystals rapidly reduced primary human chondrocyte and cartilage explant viability in a dose-dependent manner (p gout, normal cartilage architecture was lost, with empty chondrocyte lacunae observed. MSU crystals have profound inhibitory effects on chondrocyte viability and function. Interactions between MSU crystals and chondrocytes may contribute to cartilage damage in gout through reduction of chondrocyte viability and promotion of a catabolic state.

  13. Autologous chondrocyte implantation: Is it likely to become a saviour of large-sized and full-thickness cartilage defect in young adult knee?

    Science.gov (United States)

    Zhang, Chi; Cai, You-Zhi; Lin, Xiang-Jin

    2016-05-01

    A literature review of the first-, second- and third-generation autologous chondrocyte implantation (ACI) technique for the treatment of large-sized (>4 cm(2)) and full-thickness knee cartilage defects in young adults was conducted, examining the current literature on features, clinical scores, complications, magnetic resonance image (MRI) and histological outcomes, rehabilitation and cost-effectiveness. A literature review was carried out in the main medical databases to evaluate the several studies concerning ACI treatment of large-sized and full-thickness knee cartilage defects in young adults. ACI technique has been shown to relieve symptoms and improve functional assessment in large-sized (>4 cm(2)) and full-thickness knee articular cartilage defect of young adults in short- and medium-term follow-up. Besides, low level of evidence demonstrated its efficiency and durability at long-term follow-up after implantation. Furthermore, MRI and histological evaluations provided the evidence that graft can return back to the previous nearly normal cartilage via ACI techniques. Clinical outcomes tend to be similar in different ACI techniques, but with simplified procedure, low complication rate and better graft quality in the third-generation ACI technique. ACI based on the experience of cell-based therapy, with the high potential to regenerate hyaline-like tissue, represents clinical development in treatment of large-sized and full-thickness knee cartilage defects. IV.

  14. Activation of Indian Hedgehog Promotes Chondrocyte Hypertrophy and Upregulation of MMP-13 in Human Osteoarthritic Cartilage

    Science.gov (United States)

    Wei, Fangyuan; Zhou, Jingming; Wei, Xiaochun; Zhang, Juntao; Fleming, Braden C.; Terek, Richard; Pei, Ming; Chen, Qian; Liu, Tao; Wei, Lei

    2012-01-01

    Objective The objectives of this study were to 1) determine the correlation between osteoarthritis (OA) and Ihh expression, and 2) establish the effects of Ihh on expression of markers of chondrocyte hypertrophy and MMP-13 in human OA cartilage. Design OA cartilage and synovial fluid samples were obtained during total knee arthroplasty. Normal cartilage samples were obtained from intra-articular tumor resections, and normal synovial fluid samples were obtained from healthy volunteers and the contralateral uninjured knee of patients undergoing anterior cruciate ligament reconstruction. OA was graded using the Mankin score. Expression of Ihh in synovial fluid was determined by western blot. Ihh, type X collagen and MMP-13 mRNA were determined by real time PCR. Protein expression of type X collagen and MMP-13 in cartilage samples were analyzed with immunohistochemistry. Chondrocyte size was measured using image analysis. Results Ihh expression was increased 2.6 fold in OA cartilage and 37% in OA synovial fluid when compared to normal control samples. Increased expression of Ihh was associated with the severity of OA and expression of markers of chondrocyte hypertrophy: type X collagen and MMP-13, and chondocyte size. Chondrocytes were more spherical with increasing severity of OA. There was a significant correlation between Mankin score and cell size (r2= 0.80) and Ihh intensity (r2 = 0.89). Exogenous Ihh induced a 6.8 fold increase of type X collagen and 2.8 fold increase of MMP-13 mRNA expression in cultured chondrocytes. Conversely, knockdown of Ihh by siRNA and Hh inhibitor Cyclopamine had the opposite effect. Conclusions Ihh expression correlates with OA progression and changes in chondrocyte morphology and gene expression consistent with chondrocyte hypertrophy and cartilage degradation seen in OA cartilage. Thus, Ihh may be a potential therapeutic target to prevent OA progression. PMID:22469853

  15. Investigation of the Effects of Extracellular Osmotic Pressure on Morphology and Mechanical Properties of Individual Chondrocyte.

    Science.gov (United States)

    Nguyen, Trung Dung; Oloyede, Adekunle; Singh, Sanjleena; Gu, YuanTong

    2016-06-01

    It has been demonstrated that most cells of the body respond to osmotic pressure in a systematic manner. The disruption of the collagen network in the early stages of osteoarthritis causes an increase in water content of cartilage which leads to a reduction of pericellular osmolality in chondrocytes distributed within the extracellular environment. It is therefore arguable that an insight into the mechanical properties of chondrocytes under varying osmotic pressure would provide a better understanding of chondrocyte mechanotransduction and potentially contribute to knowledge on cartilage degeneration. In this present study, the chondrocyte cells were exposed to solutions with different osmolality. Changes in their dimensions and mechanical properties were measured over time. Atomic force microscopy (AFM) was used to apply load at various strain-rates and the force-time curves were logged. The thin-layer elastic model was used to extract the elastic stiffness of chondrocytes at different strain-rates and at different solution osmolality. In addition, the porohyperelastic (PHE) model was used to investigate the strain-rate-dependent responses under the loading and osmotic pressure conditions. The results revealed that the hypo-osmotic external environment increased chondrocyte dimensions and reduced Young's modulus of the cells at all strain-rates tested. In contrast, the hyper-osmotic external environment reduced dimensions and increased Young's modulus. Moreover, using the PHE model coupled with inverse FEA simulation, we established that the hydraulic permeability of chondrocytes increased with decreasing extracellular osmolality which is consistent with previous work in the literature. This could be due to a higher intracellular fluid volume fraction with lower osmolality.

  16. Study of differential properties of fibrochondrocytes and hyaline chondrocytes in growing rabbits.

    Science.gov (United States)

    Huang, L; Li, M; Li, H; Yang, C; Cai, X

    2015-02-01

    We aimed to build a culture model of chondrocytes in vitro, and to study the differential properties between fibrochondrocytes and hyaline chondrocytes. Histological sections were stained with haematoxylin and eosin so that we could analyse the histological structure of the fibrocartilage and hyaline cartilage. Condylar fibrochondrocytes and femoral hyaline chondrocytes were cultured from four, 4-week-old, New Zealand white rabbits. The production of COL2A1, COL1OA1, SOX9 and aggrecan was detected by real time-q polymerase chain reaction (RT-qPCR) and immunoblotting and the differences between them were compared statistically. Histological structures obviously differed between fibrocartilage and hyaline cartilage. COL2A1 and SOX9 were highly expressed within cell passage 2 (P2) of both fibrochondrocytes and hyaline chondrocytes, and reduced significantly after cell passage 4 (P4). The mRNA expressions of COL2A1 (p=0.05), COL10A1 (p=0.04), SOX9 (p=0.03), and aggrecan (p=0.04) were significantly higher in hyaline chondrocytes than in fibrochondrocytes, whereas the expression of COL1A1 (p=0.02) was the opposite. Immunoblotting showed similar results. We have built a simple and effective culture model of chondrocytes in vitro, and the P2 of chondrocytes is recommended for further studies. Condylar fibrocartilage and femoral hyaline cartilage have unique biological properties, and the regulatory mechanisms of endochondral ossification for the condyle should be studied independently in the future. Copyright © 2014 The British Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

  17. Primary chondrocytes enhance cartilage tissue formation upon co-culture with expanded chondrocytes, dermal fibroblasts, 3T3 feeder cells and embryonic stem cells

    NARCIS (Netherlands)

    Hendriks, J.A.A.; Miclea, Razvan L.; Schotel, Roka; de Bruijn, Ewart; Moroni, Lorenzo; Karperien, Hermanus Bernardus Johannes; Riesle, J.U.; van Blitterswijk, Clemens

    2010-01-01

    Co-culture models have been increasingly used in tissue engineering applications to understand cell–cell interactions and consequently improve regenerative medicine strategies. Aiming at further elucidating cartilage tissue formation, we co-cultured bovine primary chondrocytes (BPCs) with human

  18. Mathematical modelling of tissue formation in chondrocyte filter cultures.

    Science.gov (United States)

    Catt, C J; Schuurman, W; Sengers, B G; van Weeren, P R; Dhert, W J A; Please, C P; Malda, J

    2011-12-17

    In the field of cartilage tissue engineering, filter cultures are a frequently used three-dimensional differentiation model. However, understanding of the governing processes of in vitro growth and development of tissue in these models is limited. Therefore, this study aimed to further characterise these processes by means of an approach combining both experimental and applied mathematical methods. A mathematical model was constructed, consisting of partial differential equations predicting the distribution of cells and glycosaminoglycans (GAGs), as well as the overall thickness of the tissue. Experimental data was collected to allow comparison with the predictions of the simulation and refinement of the initial models. Healthy mature equine chondrocytes were expanded and subsequently seeded on collagen-coated filters and cultured for up to 7 weeks. Resulting samples were characterised biochemically, as well as histologically. The simulations showed a good representation of the experimentally obtained cell and matrix distribution within the cultures. The mathematical results indicate that the experimental GAG and cell distribution is critically dependent on the rate at which the cell differentiation process takes place, which has important implications for interpreting experimental results. This study demonstrates that large regions of the tissue are inactive in terms of proliferation and growth of the layer. In particular, this would imply that higher seeding densities will not significantly affect the growth rate. A simple mathematical model was developed to predict the observed experimental data and enable interpretation of the principal underlying mechanisms controlling growth-related changes in tissue composition.

  19. From gristle to chondrocyte transplantation: treatment of cartilage injuries.

    Science.gov (United States)

    Lindahl, Anders

    2015-10-19

    This review addresses the progress in cartilage repair technology over the decades with an emphasis on cartilage regeneration with cell therapy. The most abundant cartilage is the hyaline cartilage that covers the surface of our joints and, due to avascularity, this tissue is unable to repair itself. The cartilage degeneration seen in osteoarthritis causes patient suffering and is a huge burden to society. The surgical approach to cartilage repair was non-existing until the 1950s when new surgical techniques emerged. The use of cultured cells for cell therapy started as experimental studies in the 1970s that developed over the years to a clinical application in 1994 with the introduction of the autologous chondrocyte transplantation technique (ACT). The technology is now spread worldwide and has been further refined by combining arthroscopic techniques with cells cultured on matrix (MACI technology). The non-regenerating hypothesis of cartilage has been revisited and we are now able to demonstrate cell divisions and presence of stem-cell niches in the joint. Furthermore, cartilage derived from human embryonic stem cells and induced pluripotent stem cells could be the base for new broader cell treatments for cartilage injuries and the future technology base for prevention and cure of osteoarthritis. © 2015 The Author(s).

  20. From gristle to chondrocyte transplantation: treatment of cartilage injuries

    Science.gov (United States)

    Lindahl, Anders

    2015-01-01

    This review addresses the progress in cartilage repair technology over the decades with an emphasis on cartilage regeneration with cell therapy. The most abundant cartilage is the hyaline cartilage that covers the surface of our joints and, due to avascularity, this tissue is unable to repair itself. The cartilage degeneration seen in osteoarthritis causes patient suffering and is a huge burden to society. The surgical approach to cartilage repair was non-existing until the 1950s when new surgical techniques emerged. The use of cultured cells for cell therapy started as experimental studies in the 1970s that developed over the years to a clinical application in 1994 with the introduction of the autologous chondrocyte transplantation technique (ACT). The technology is now spread worldwide and has been further refined by combining arthroscopic techniques with cells cultured on matrix (MACI technology). The non-regenerating hypothesis of cartilage has been revisited and we are now able to demonstrate cell divisions and presence of stem-cell niches in the joint. Furthermore, cartilage derived from human embryonic stem cells and induced pluripotent stem cells could be the base for new broader cell treatments for cartilage injuries and the future technology base for prevention and cure of osteoarthritis. PMID:26416680

  1. Vein grafting in fingertip replantations.

    Science.gov (United States)

    Yan, Hede; Jackson, William D; Songcharoen, Somjade; Akdemir, Ovunc; Li, Zhijie; Chen, Xinglong; Jiang, Liangfu; Gao, Weiyang

    2009-01-01

    In this retrospective study, the survival rates of fingertip replantation with and without vein grafting were evaluated along with their postoperative functional and cosmetic results. One hundred twenty-one-fingertip amputations were performed in 103 patients between September 2002 and July 2007. Thirty-four amputated fingertips were replanted without vein grafting, while 87 amputated fingertips were replanted with vein grafting for arterial and/or venous repairs. The overall survival rates of the replantations with and without vein grafting were 90% (78/87) and 85% (29/34), respectively. The survival rates were 88% (36/41) with venous repair, 93% (25/27) with arterial repair, and 89% (17/19) with both. Nineteen patients without vein grafting and 48 patients with vein grafting had a follow-up period of more than one year. Good cosmetic and functional outcomes were observed in both groups of patients. The results show that vein grafting is a reliable technique in fingertip replantations, showing no significant difference (P > 0.05) in survival between those with and without vein grafting. Furthermore, no significant difference (P > 0.05) in survival was found between cases with vein grafts for arterial and/or venous repairs. In fingertip replantations with vein grafting, favorable functional and esthetic results can be achieved without sacrificing replantation survival. (c) 2009 Wiley-Liss, Inc.

  2. A novel fibrin gel derived from hyaluronic acid-grafted fibrinogen

    International Nuclear Information System (INIS)

    Yang, Chiung L; Chen, Hui W; Wang, Tzu C; Wang, Yng J

    2011-01-01

    Fibrinogen is a major plasma protein that forms a three-dimensional fibrin gel upon being activated by thrombin. In this study, we report the synthesis and potential applications of hybrid molecules composed of fibrinogen coupled to the reducing ends of short-chain hyaluronic acids (sHAs) by reductive amination. The grafting of sHAs to fibrinogen was verified by analyzing particle size, zeta potential and gel-electrophoretic mobility of the hybrid molecules. The sHA-fibrinogen hybrid molecules with graft ratios (sHA/fibrinogen) of up to 6.5 retained the ability to form gels in response to thrombin activation. The sHA-fibrin gels were transparent in appearance and exhibited high water content, which were characteristics distinct from those of gels formed by mixtures of sHAs and fibrinogen. The potential applications of the sHA-fibrin gels were evaluated. The sHA-fibrinogen gel with a graft ratio of 3.6 (S3.6F) was examined for its ability to encapsulate and support the differentiation of ATDC5 chondrocyte-like cells. Compared with the fibrinogen-formed gel, cells cultured in the S3.6F gel exhibited increased lacunae formation; moreover, the abundance of cartilaginous extracellular matrix molecules and the expression of chondrocyte marker genes, such as aggrecan, collagen II and Sox9, were also significantly increased. Our data suggest that the three-dimensional gel formed by the sHA-fibrinogen hybrid is a better support than the fibrin gel for chondrogenesis induction.

  3. A novel fibrin gel derived from hyaluronic acid-grafted fibrinogen

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Chiung L; Chen, Hui W; Wang, Tzu C; Wang, Yng J, E-mail: wang@ym.edu.tw [Institute of Biomedical Engineering, National Yang Ming University, No. 155, Sec. 2, Li-Nung St., Shih-Pai, Taipei, Taiwan 112 (China)

    2011-04-15

    Fibrinogen is a major plasma protein that forms a three-dimensional fibrin gel upon being activated by thrombin. In this study, we report the synthesis and potential applications of hybrid molecules composed of fibrinogen coupled to the reducing ends of short-chain hyaluronic acids (sHAs) by reductive amination. The grafting of sHAs to fibrinogen was verified by analyzing particle size, zeta potential and gel-electrophoretic mobility of the hybrid molecules. The sHA-fibrinogen hybrid molecules with graft ratios (sHA/fibrinogen) of up to 6.5 retained the ability to form gels in response to thrombin activation. The sHA-fibrin gels were transparent in appearance and exhibited high water content, which were characteristics distinct from those of gels formed by mixtures of sHAs and fibrinogen. The potential applications of the sHA-fibrin gels were evaluated. The sHA-fibrinogen gel with a graft ratio of 3.6 (S3.6F) was examined for its ability to encapsulate and support the differentiation of ATDC5 chondrocyte-like cells. Compared with the fibrinogen-formed gel, cells cultured in the S3.6F gel exhibited increased lacunae formation; moreover, the abundance of cartilaginous extracellular matrix molecules and the expression of chondrocyte marker genes, such as aggrecan, collagen II and Sox9, were also significantly increased. Our data suggest that the three-dimensional gel formed by the sHA-fibrinogen hybrid is a better support than the fibrin gel for chondrogenesis induction.

  4. Downregulation of protein kinase CK2 activity facilitates tumor necrosis factor-α-mediated chondrocyte death through apoptosis and autophagy.

    Directory of Open Access Journals (Sweden)

    Sung Won Lee

    Full Text Available Despite the numerous studies of protein kinase CK2, little progress has been made in understanding its function in chondrocyte death. Our previous study first demonstrated that CK2 is involved in apoptosis of rat articular chondrocytes. Recent studies have suggested that CK2 downregulation is associated with aging. Thus examining the involvement of CK2 downregulation in chondrocyte death is an urgently required task. We undertook this study to examine whether CK2 downregulation modulates chondrocyte death. We first measured CK2 activity in articular chondrocytes of 6-, 21- and 30-month-old rats. Noticeably, CK2 activity was downregulated in chondrocytes with advancing age. To build an in vitro experimental system for simulating tumor necrosis factor (TNF-α-induced cell death in aged chondrocytes with decreased CK2 activity, chondrocytes were co-treated with CK2 inhibitors and TNF-α. Viability assay demonstrated that CK2 inhibitors facilitated TNF-α-mediated chondrocyte death. Pulsed-field gel electrophoresis, nuclear staining, flow cytometry, TUNEL staining, confocal microscopy, western blot and transmission electron microscopy were conducted to assess cell death modes. The results of multiple assays showed that this cell death was mediated by apoptosis. Importantly, autophagy was also involved in this process, as supported by the appearance of a punctuate LC3 pattern and autophagic vacuoles. The inhibition of autophagy by silencing of autophage-related genes 5 and 7 as well as by 3-methyladenine treatment protected chondrocytes against cell death and caspase activation, indicating that autophagy led to the induction of apoptosis. Autophagic cells were observed in cartilage obtained from osteoarthritis (OA model rats and human OA patients. Our findings indicate that CK2 down regulation facilitates TNF-α-mediated chondrocyte death through apoptosis and autophagy. It should be clarified in the future if autophagy observed is a consequence

  5. Autologous Chondrocyte Implantation for Bipolar Chondral Lesions in the Tibiofemoral Compartment.

    Science.gov (United States)

    Ogura, Takahiro; Bryant, Tim; Mosier, Brian A; Minas, Tom

    2018-05-01

    Treating bipolar chondral lesions in the tibiofemoral (TF) compartment with cartilage repair procedures is challenging, and a suitable treatment remains unclear. To evaluate clinical outcomes after autologous chondrocyte implantation (ACI) for the treatment of bipolar chondral lesions in the TF compartment. Case series; Level of evidence, 4. We evaluated 57 patients who underwent ACI for the treatment of symptomatic bipolar chondral lesions in the TF compartment by a single surgeon between October 1995 and June 2014. One patient did not return for follow-up. Thus, 56 patients (58 knees) were included with a minimum of 2 years' follow-up. A mean of 3.1 lesions per knee were treated, representing a mean total surface area of 16.1 cm 2 (range, 3.2-44.5 cm 2 ) per knee. Bipolar lesions were present in the medial compartment (32 knees) and in the lateral compartment (26 knees). Patients were evaluated with the modified Cincinnati Knee Rating Scale, visual analog scale for pain, Western Ontario and McMaster Universities Osteoarthritis Index, and Short Form-36. Patients also answered questions regarding self-rated knee function and satisfaction with the procedure. Standard radiographs were evaluated with the Kellgren-Lawrence grading system. The survival rate was 80% at 5 years and 76% at 10 years. A significantly better survival rate was found in patients with the use of a collagen membrane than periosteum (97% vs 61% at 5 years, respectively; P = .0014). Of 46 knees with retained grafts, all functional scores significantly improved postoperatively, with a very high satisfaction rate (91%) at a mean of 8.3 ± 5.1 years (range, 2-20 years) after ACI. At last follow-up, 24 of 46 successful knees were radiographically assessed (mean, 5.5 ± 4.0 years [range, 2.0-18.7 years]) and showed no significant osteoarthritis progression ( P = .3173). Outcomes for 12 patients were considered as failures at a mean of 4.1 years. Of these, 9 patients were converted to partial or total

  6. Mechanical and hypoxia stress can cause chondrocytes apoptosis through over-activation of endoplasmic reticulum stress.

    Science.gov (United States)

    Huang, Ziwei; Zhou, Min; Wang, Qian; Zhu, Mengjiao; Chen, Sheng; Li, Huang

    2017-12-01

    To examine the role of mechanical force and hypoxia on chondrocytes apoptosis and osteoarthritis (OA)-liked pathological change on mandibular cartilage through over-activation of endoplasmic reticulum stress (ERS). We used two in vitro models to examine the effect of mechanical force and hypoxia on chondrocytes apoptosis separately. The mandibular condylar chondrocytes were obtained from three-week-old male Sprague-Dawley rats. Flexcell 5000T apparatus was used to produce mechanical forces (12%, 0.5Hz, 24h vs 20%, 0.5Hz, 24h) on chondrocytes. For hypoxia experiment, the concentration of O 2 was down regulated to 5% or 1%. Cell apoptosis rates were quantified by annexin V and propidium iodide (PI) double staining and FACS analysis. Quantitative real-time PCR and western blot were performed to evaluate the activation of ERS and cellular hypoxia. Then we used a mechanical stress loading rat model to verify the involvement of ERS in OA-liked mandibular cartilage pathological change. Histological changes in mandibular condylar cartilage were assessed via hematoxylin & eosin (HE) staining. Immunohistochemistry of GRP78, GRP94, HIF-1α, and HIF-2α were performed to evaluate activation of the ERS and existence of hypoxia. Apoptotic cells were detected by the TUNEL method. Tunicamycin, 20% mechanical forces and hypoxia (1% O 2 ) all significantly increased chondrocytes apoptosis rates and expression of ERS markers (GRP78, GRP94 and Caspase 12). However, 12% mechanical forces can only increase the apoptotic sensitivity of chondrocytes. Mechanical stress resulted in OA-liked pathological change on rat mandibular condylar cartilage which included thinning cartilage and bone erosion. The number of apoptotic cells increased. ERS and hypoxia markers expressions were also enhanced. Salubrinal, an ERS inhibitor, can reverse these effects in vitro and in vivo through the down-regulation of ERS markers and hypoxia markers. We confirmed that mechanical stress and local hypoxia both

  7. One-stage focal cartilage defect treatment with bone marrow mononuclear cells and chondrocytes leads to better macroscopic cartilage regeneration compared to microfracture in goats

    NARCIS (Netherlands)

    Bekkers, J.E.J.; Creemers, L.B.; Tsuchida, A.I.; van Rijen, M.H.P.; Custers, R.J.H.; Dhert, W.J.A.; Saris, Daniël B.F.

    2013-01-01

    Objective The combination of chondrocytes and mononuclear fraction (MNF) cells might solve the expansion induced dedifferentiation problem of reimplanted cells in autologous chondrocytes implantation as sufficient cells would be available for direct, one-stage, implantation. Earlier in vitro work

  8. The Regulatory Role of Signaling Crosstalk in Hypertrophy of MSCs and Human Articular Chondrocytes

    Directory of Open Access Journals (Sweden)

    Leilei Zhong

    2015-08-01

    Full Text Available Hypertrophic differentiation of chondrocytes is a main barrier in application of mesenchymal stem cells (MSCs for cartilage repair. In addition, hypertrophy occurs occasionally in osteoarthritis (OA. Here we provide a comprehensive review on recent literature describing signal pathways in the hypertrophy of MSCs-derived in vitro differentiated chondrocytes and chondrocytes, with an emphasis on the crosstalk between these pathways. Insight into the exact regulation of hypertrophy by the signaling network is necessary for the efficient application of MSCs for articular cartilage repair and for developing novel strategies for curing OA. We focus on articles describing the role of the main signaling pathways in regulating chondrocyte hypertrophy-like changes. Most studies report hypertrophic differentiation in chondrogenesis of MSCs, in both human OA and experimental OA. Chondrocyte hypertrophy is not under the strict control of a single pathway but appears to be regulated by an intricately regulated network of multiple signaling pathways, such as WNT, Bone morphogenetic protein (BMP/Transforming growth factor-β (TGFβ, Parathyroid hormone-related peptide (PTHrP, Indian hedgehog (IHH, Fibroblast growth factor (FGF, Insulin like growth factor (IGF and Hypoxia-inducible factor (HIF. This comprehensive review describes how this intricate signaling network influences tissue-engineering applications of MSCs in articular cartilage (AC repair, and improves understanding of the disease stages and cellular responses within an OA articular joint.

  9. Subchondral Bone Plate Thickening Precedes Chondrocyte Apoptosis and Cartilage Degradation in Spontaneous Animal Models of Osteoarthritis

    Directory of Open Access Journals (Sweden)

    Zaitunnatakhin Zamli

    2014-01-01

    Full Text Available Osteoarthritis (OA is the most common joint disorder characterised by bone remodelling and cartilage degradation and associated with chondrocyte apoptosis. These processes were investigated at 10, 16, 24, and 30 weeks in Dunkin Hartley (DH and Bristol Strain 2 (BS2 guinea pigs that develop OA spontaneously. Both strains had a more pronounced chondrocyte apoptosis, cartilage degradation, and subchondral bone changes in the medial than the lateral side of the tibia, and between strains, the changes were always greater and faster in DH than BS2. In the medial side, a significant increase of chondrocyte apoptosis and cartilage degradation was observed in DH between 24 and 30 weeks of age preceded by a progressive thickening and stiffening of subchondral bone plate (Sbp. The Sbp thickness consistently increased over the 30-week study period but the bone mineral density (BMD of the Sbp gradually decreased after 16 weeks. The absence of these changes in the medial side of BS2 may indicate that the Sbp of DH was undergoing remodelling. Chondrocyte apoptosis was largely confined to the deep zone of articular cartilage and correlated with thickness of the subchondral bone plate suggesting that cartilage degradation and chondrocyte apoptosis may be a consequence of continuous bone remodelling during the development of OA in these animal models of OA.

  10. Treatment of osteoarthritis using a helper-dependent adenoviral vector retargeted to chondrocytes

    Directory of Open Access Journals (Sweden)

    Merry ZC Ruan

    2016-01-01

    Full Text Available Osteoarthritis (OA is a joint disease characterized by degeneration of the articular cartilage, subchondral bone remodeling, and secondary inflammation. It is among the top three causes of chronic disability, and currently there are no treatment options to prevent disease progression. The localized nature of OA makes it an ideal candidate for gene and cell therapy. However, gene and cell therapy of OA is impeded by inefficient gene transduction of chondrocytes. In this study, we developed a broadly applicable system that retargets cell surface receptors by conjugating antibodies to the capsid of helper-dependent adenoviral vectors (HDVs. Specifically, we applied this system to retarget chondrocytes by conjugating an HDV to an α-10 integrin monoclonal antibody (a10mab. We show that a10mab-conjugated HDV (a10mabHDV-infected chondrocytes efficiently in vitro and in vivo while detargeting other cell types. The therapeutic index of an intra-articular injection of 10mabHDV-expressing proteoglycan 4 (PRG4 into a murine model of post-traumatic OA was 10-fold higher than with standard HDV. Moreover, we show that PRG4 overexpression from articular, superficial zone chondrocytes is effective for chondroprotection in postinjury OA and that α-10 integrin is an effective protein for chondrocyte targeting.

  11. Normal proliferation and differentiation of Hoxc-8 transgenic chondrocytes in vitro

    Directory of Open Access Journals (Sweden)

    Mello Maria

    2003-04-01

    Full Text Available Abstract Background Hox genes encode transcription factors that are involved in pattern formation in the skeleton, and recent evidence suggests that they also play a role in the regulation of endochondral ossification. To analyze the role of Hoxc-8 in this process in more detail, we applied in vitro culture systems, using high density cultures of primary chondrocytes from neonatal mouse ribs. Results Cultured cells were characterized on the basis of morphology (light microscopy and production of cartilage-specific extracellular matrix (sulfated proteoglycans and type II Collagen. Hypertrophy was demonstrated by increase in cell size, alkaline phosphatase activity and type X Collagen immunohistochemistry. Proliferation was assessed by BrdU uptake and flow cytometry. Unexpectedly, chondrocytes from Hoxc-8 transgenic mice, which exhibit delayed cartilage maturation in vivo 1, were able to proliferate and differentiate normally in our culture systems. This was the case even though freshly isolated Hoxc-8 transgenic chondrocytes exhibited significant molecular differences as measured by real-time quantitative PCR. Conclusions The results demonstrate that primary rib chondrocytes behave similar to published reports for chondrocytes from other sources, validating in vitro approaches for studies of Hox genes in the regulation of endochondral ossification. Our analysis of cartilage-producing cells from Hoxc-8 transgenic mice provides evidence that the cellular phenotype induced by Hoxc-8 overexpression in vivo is reversible in vitro.

  12. RhoA activation and nuclearization marks loss of chondrocyte phenotype in crosstalk with Wnt pathway.

    Science.gov (United States)

    Öztürk, Ece; Despot-Slade, Evelin; Pichler, Michael; Zenobi-Wong, Marcy

    2017-11-15

    De-differentiation comprises a major drawback for the use of autologous chondrocytes in cartilage repair. Here, we investigate the role of RhoA and canonical Wnt signaling in chondrocyte phenotype. Chondrocyte de-differentiation is accompanied by an upregulation and nuclear localization of RhoA. Effectors of canonical Wnt signaling including β-catenin and YAP/TAZ are upregulated in de-differentiating chondrocytes in a Rho-dependent manner. Inhibition of Rho activation with C3 transferase inhibits nuclear localization of RhoA, induces expression of chondrogenic markers on 2D and enhances the chondrogenic effect of 3D culturing. Upregulation of chondrogenic markers by Rho inhibition is accompanied by loss of canonical Wnt signaling markers in 3D or on 2D whereas treatment of chondrocytes with Wnt-3a abrogates this effect. However, induction of canonical Wnt signaling inhibits chondrogenic markers on 2D but enhances chondrogenic re-differentiation on 2D with C3 transferase or in 3D. These data provide insights on the context-dependent role of RhoA and Wnt signaling in de-differentiation and on mechanisms to induce chondrogenic markers for therapeutic approaches. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Single Cell Confocal Raman Spectroscopy of Human Osteoarthritic Chondrocytes: A Preliminary Study

    Directory of Open Access Journals (Sweden)

    Rajesh Kumar

    2015-04-01

    Full Text Available A great deal of effort has been focused on exploring the underlying molecular mechanism of osteoarthritis (OA especially at the cellular level. We report a confocal Raman spectroscopic investigation on human osteoarthritic chondrocytes. The objective of this investigation is to identify molecular features and the stage of OA based on the spectral signatures corresponding to bio-molecular changes at the cellular level in chondrocytes. In this study, we isolated chondrocytes from human osteoarthritic cartilage and acquired Raman spectra from single cells. Major spectral differences between the cells obtained from different International Cartilage Repair Society (ICRS grades of osteoarthritic cartilage were identified. During progression of OA, a decrease in protein content and an increase in cell death were observed from the vibrational spectra. Principal component analysis and subsequent cross-validation was able to associate osteoarthritic chondrocytes to ICRS Grade I, II and III with specificity 100.0%, 98.1%, and 90.7% respectively, while, sensitivity was 98.6%, 82.8%, and 97.5% respectively. The overall predictive efficiency was 92.2%. Our pilot study encourages further use of Raman spectroscopy as a noninvasive and label free technique for revealing molecular features associated with osteoarthritic chondrocytes.

  14. Cellular automata model for human articular chondrocytes migration, proliferation and cell death: An in vitro validation.

    Science.gov (United States)

    Vaca-González, J J; Gutiérrez, M L; Guevara, J M; Garzón-Alvarado, D A

    2017-01-01

    Articular cartilage is characterized by low cell density of only one cell type, chondrocytes, and has limited self-healing properties. When articular cartilage is affected by traumatic injuries, a therapeutic strategy such as autologous chondrocyte implantation is usually proposed for its treatment. This approach requires in vitro chondrocyte expansion to yield high cell number for cell transplantation. To improve the efficiency of this procedure, it is necessary to assess cell dynamics such as migration, proliferation and cell death during culture. Computational models such as cellular automata can be used to simulate cell dynamics in order to enhance the result of cell culture procedures. This methodology has been implemented for several cell types; however, an experimental validation is required for each one. For this reason, in this research a cellular automata model, based on random-walk theory, was devised in order to predict articular chondrocyte behavior in monolayer culture during cell expansion. Results demonstrated that the cellular automata model corresponded to cell dynamics and computed-accurate quantitative results. Moreover, it was possible to observe that cell dynamics depend on weighted probabilities derived from experimental data and cell behavior varies according to the cell culture period. Thus, depending on whether cells were just seeded or proliferated exponentially, culture time probabilities differed in percentages in the CA model. Furthermore, in the experimental assessment a decreased chondrocyte proliferation was observed along with increased passage number. This approach is expected to having other uses as in enhancing articular cartilage therapies based on tissue engineering and regenerative medicine.

  15. Effects of tofacitinib on nucleic acid metabolism in human articular chondrocytes.

    Science.gov (United States)

    Koizumi, Hideki; Arito, Mitsumi; Endo, Wataru; Kurokawa, Manae S; Okamoto, Kazuki; Omoteyama, Kazuki; Suematsu, Naoya; Beppu, Moroe; Kato, Tomohiro

    2015-07-01

    In our previous screening of chondrocyte protein profiles, the amount of adenosine monophosphate deaminase (AMPD) 2 was found to be decreased by tofacitinib. Extending the study, here we confirmed the decrease of AMPD2 by tofacitinib and further investigated effects of tofacitinib on purine nucleotide metabolism. Human articular chondrocytes and a chondrosarcoma cell line: OUMS-27 were stimulated with tofacitinib. Then the levels of AMPD2 and its related enzymes were investigated by Western blot. The levels of AMP and adenosine were assessed by mass spectrometry. We confirmed the significant decrease of AMPD2 by tofacitinib in chondrocytes (p = 0.025). The levels of adenosine kinase and 5'-nucleotidase were decreased in chondrocytes, although they did not meet statistical significance (p = 0.067 and p = 0.074, respectively). The results from OUMS-27 were similar to those from the chondrocytes. The cellular adenosine levels were significantly decreased by tofacitinib in OUMS-27 (p = 0.014). The cellular AMP levels were increased, although they did not meet statistical significance in OUMS-27 (p = 0.066). Our data indicate that tofacitinib increases the cellular levels of adenosine, which is known to have anti-inflammatory activity, through the downregulation of AMPD2. This would be a novel functional aspect of tofacitinib.

  16. Comparing effects of perfusion and hydrostatic pressure on gene profiles of human chondrocyte.

    Science.gov (United States)

    Zhu, Ge; Mayer-Wagner, Susanne; Schröder, Christian; Woiczinski, Matthias; Blum, Helmut; Lavagi, Ilaria; Krebs, Stefan; Redeker, Julia I; Hölzer, Andreas; Jansson, Volkmar; Betz, Oliver; Müller, Peter E

    2015-09-20

    Hydrostatic pressure and perfusion have been shown to regulate the chondrogenic potential of articular chondrocytes. In order to compare the effects of hydrostatic pressure plus perfusion (HPP) and perfusion (P) we investigated the complete gene expression profiles of human chondrocytes under HPP and P. A simplified bioreactor was constructed to apply loading (0.1 MPa for 2 h) and perfusion (2 ml) through the same piping by pressurizing the medium directly. High-density monolayer cultures of human chondrocytes were exposed to HPP or P for 4 days. Controls (C) were maintained in static cultures. Gene expression was evaluated by sequencing (RNAseq) and quantitative real-time PCR analysis. Both treatments changed gene expression levels of human chondrocytes significantly. Specifically, HPP and P increased COL2A1 expression and decreased COL1A1 and MMP-13 expression. Despite of these similarities, RNAseq revealed a list of cartilage genes including ACAN, ITGA10 and TNC, which were differentially expressed by HPP and P. Of these candidates, adhesion related molecules were found to be upregulated in HPP. Both HPP and P treatment had beneficial effects on chondrocyte differentiation and decreased catabolic enzyme expression. The study provides new insight into how hydrostatic pressure and perfusion enhance cartilage differentiation and inhibit catabolic effects. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Analysis of human articular chondrocyte CD44 isoform expression and function in health and disease.

    Science.gov (United States)

    Salter, D M; Godolphin, J L; Gourlay, M S; Lawson, M F; Hughes, D E; Dunne, E

    1996-08-01

    Interactions between articular chondrocytes and components of the extracellular matrix are of potential importance in the normal function of cartilage and in the pathophysiology of arthritis. Little is known of the basis of these interactions, but cell adhesive molecules such as CD44 are likely to be involved. Immunohistology using six well-characterized anti-CD44 monoclonal antibodies demonstrated standard CD44 isoform (CD44H) expression by all chondrocytes in normal and osteoarthrotic (OA) cartilage but absence of the CD44E variant. Polymerase chain reaction (PCR) of reverse transcribed mRNA from monolayer cultures of normal and OA chondrocytes using primer sequences which span the region containing variably spliced exons produced a predominant band representing the standard form of CD44, which lacks the variable exons 6-15 (v1-v10). No product was seen at the expected size of the epithelial variant of CD44 (CD44v8-10). Use of exon-specific primers, however, showed expression of variant exons resulting in multiple minor isoforms. Standard CD44 was also shown to be the predominantly expressed isoform identified by immunoprecipitation, but human articular chondrocytes did not adhere to hyaluronan in vitro. Chondrocyte CD44 may function as an adhesion receptor for other matrix molecules such as fibronectin or collagen.

  18. Post-traumatic stress disorder and its predictors among bereaved Tibetan adolescents four years after the Yushu earthquake: a cross-sectional survey in China.

    Science.gov (United States)

    Dongling, Liu; Hui, Chen; Ling, Ma; Wenqian, Bie; Zailiang, Liu; Changying, Chen

    2017-04-01

    To examine the prevalence of post-traumatic stress disorder symptoms and to explore the predictive factors of post-traumatic stress disorder symptoms among bereaved adolescents four years after the Yushu earthquake. On 14 April 2010, the 7·1-magnitude Yushu earthquake occurred in the high-altitude, hypoxia-prone regions primarily inhabited by ethnic minorities. Many adolescents lost their parents during the earthquake. This study examined post-traumatic stress disorder and its predictors among bereaved Tibetan adolescents four years after the trauma in China. This was a cross-sectional descriptive study. This study used a cross-sectional design with 830 bereaved adolescents. Participant demographic data included gender, age and grade, and the earthquake exposure variables included the deceased relatives, whether they were buried, injured or amputated (non-life-threatening) and whether they witnessed a burial, injury or death. The post-traumatic stress disorder Checklist-Civilian version was used to assess the symptoms and prevalence of post-traumatic stress disorder, and the Coping Style Scale assessed coping styles. The results indicated that the prevalence of post-traumatic stress disorder was 19·3%. Individuals who were buried, injured or amputated (non-life-threatening), who witnessed a burial, injury or death, who suffered severe property loss during the earthquake and who had negative coping skills were the most likely individuals to have post-traumatic stress disorder symptoms. The presence of post-traumatic stress disorder was very prevalent among adolescents four years after the Yushu earthquake. Effective mental health services should be developed to facilitate postdisaster recovery for bereaved adolescents at high risk for post-traumatic stress disorder. The findings in this study improve our understanding of post-traumatic stress disorder and related risk factors in bereaved adolescents in non-Western communities, providing useful information for

  19. Vascular graft infections with Mycoplasma

    DEFF Research Database (Denmark)

    Levi-Mazloum, Niels Donald; Skov Jensen, J; Prag, J

    1995-01-01

    laboratory techniques, the percentage of culture-negative yet grossly infected vascular grafts seems to be increasing and is not adequately explained by the prior use of antibiotics. We have recently reported the first case of aortic graft infection with Mycoplasma. We therefore suggest the hypothesis...... that the large number of culture-negative yet grossly infected vascular grafts may be due to Mycoplasma infection not detected with conventional laboratory technique....

  20. Impact factor (if) of hospitality, leisure, sports & tourism journals: current trends, overall ranking and temporal stability over a four year period

    OpenAIRE

    Sans-Rosell, Nuria; Reverter Masià, Joaquín; Hernández González, Vicenç

    2013-01-01

    A journal’s “impact factor” (IF) is the bibliometric index that reflects the frequency with which an ‘‘average article’’ from a scientific journal has been cited in subsequent publications.The purpose of the present study is to examine the current impact factor of Hospitality, Leisure, Sports & Tourism journals, their overall ranking and temporal stability over a four year period. For this reason, we have included the impact factor of the scientific journals classifiedin the “Hospitality,...

  1. [The LORAS Project and quality assurance. In four years from input- to outcome-oriented financing in public health. 1: The LORAS Project].

    Science.gov (United States)

    Lenz, M J; Hochreutener, M A

    2001-02-01

    This series of three articles is a summary of the operations, findings and results of the hospital reform projects in the Canton of Zurich, termed LORAS. With the aid of the LORAS project within four years Zurich hospitals have been transformed. Whereas they used to adhere to input-oriented covering of deficits they now operate with outcome-oriented prospective financing of output. Part 1 describes the whole Project. Part 2 focuses on the development of outcome-measurement. Part 3 finally describes the implementation of the outcome-measurement in the canton of Zurich.

  2. [The LORAS project and quality assurance. In four years from input- to outcome-oriented financing in public health. 2: LORAS project outcome parts 1 & 98].

    Science.gov (United States)

    Lenz, M J; Hochreutener, M A

    2001-04-01

    This series of three articles is a summary of the operations, findings and results of the hospital reform projects in the Canton of Zurich, termed LORAS. With the aid of the LORAS project within four years Zurich hospitals have been transformed. Whereas they used to adhere to input-oriented covering of deficits they now operate with outcome-oriented prospective financing of output. Part 1 describes the whole project. Part 2 focuses on the development of outcome-measurement. Part 3 finally describes the implementation of the outcome-measurement in the canton of Zurich.

  3. Environmental application of radiation grafting

    International Nuclear Information System (INIS)

    Tamada, Masao

    2007-01-01

    Adsorbent having high selectivity against a certain metal ion was synthesized by means of radiation-induced graft polymerization for the purpose of environmental application. The resulting adsorbents were utilized for the removal of toxic metal from scallop waste and the collection of uranium from seawater. As a novel application of grafting, the biodegradability of poly-hydroxybutylate was controlled by grafting. The biodegradability could be depressed by the graft chain and then recovered by external stimuli such as thermal and chemical treatments. (author)

  4. Culture of chondrocytes in alginate surrounded by fibrin gel: characteristics of the cells over a period of eight weeks

    NARCIS (Netherlands)

    Almqvist, K. F.; Wang, L.; Wang, J.; Baeten, D.; Cornelissen, M.; Verdonk, R.; Veys, E. M.; Verbruggen, G.

    2001-01-01

    OBJECTIVE: To produce tissue engineered cartilage by human articular chondrocytes in vitro for further use in in vivo manipulations for the treatment of cartilage defects. METHODS: Human articular chondrocytes were cultured in 0.5%, 1.0%, and 2.0% of alginate for up to four weeks. The optimal

  5. Trophic Effects of Mesenchymal Stem Cells in Chondrocyte Co-Cultures are Independent of Culture Conditions and Cell Sources

    NARCIS (Netherlands)

    Wu, Ling; Prins, H.J.; Helder, M.; van Blitterswijk, Clemens; Karperien, Hermanus Bernardus Johannes

    2012-01-01

    Earlier, we have shown that the increased cartilage production in pellet co-cultures of chondrocytes and bone marrow-derived mesenchymal stem cells (BM-MSCs) is due to a trophic role of the MSC in stimulating chondrocyte proliferation and matrix production rather than MSCs actively undergoing

  6. Trophic effects of mesenchymal stem cells in chondrocyte co-cultures are independent of culture conditions and cell sources

    NARCIS (Netherlands)

    Wu, L.; Prins, H.J.; Helder, M.N.; van Blitterswijk, C.A.; Karperien, M.

    2012-01-01

    Earlier, we have shown that the increased cartilage production in pellet co-cultures of chondrocytes and bone marrow-derived mesenchymal stem cells (BM-MSCs) is due to a trophic role of the MSC in stimulating chondrocyte proliferation and matrix production rather than MSCs actively undergoing

  7. Comparison of Efficacy of Endogenous and Exogenous IGF-I in Stimulating Matrix Production in Neonatal and Mature Chondrocytes.

    Science.gov (United States)

    Aguilar, Izath N; Trippel, Stephen B; Shi, Shuiliang; Bonassar, Lawrence J

    2015-10-01

    The goal of this study was to compare the efficacy of endogenous upregulation of IGF-I by gene therapy and exogenous addition of insulin-like growth factor I (IGF-I) in enhancing proteoglycan synthesis by skeletally mature and neonatal chondrocytes. Chondrocyte transplantation therapy is a common treatment for focal cartilage lesions, with both mature and neonatal chondrocytes used as a cell source. Additionally, gene therapy strategies to upregulate growth factors such as IGF-I have been proposed to augment chondrocyte transplantation therapies. Both skeletally mature and neonatal chondrocytes were exposed to either an adeno-associated virus-based plasmid containing the IGF-I gene or exogenous IGF-I. Analysis of IGF-I and glycosaminoglycan production using a 4-parameter dose-response model established a clear connection between the amount of IGF-I produced by cells and their biosynthetic response. Both neonatal and mature chondrocytes showed this relationship, but the sensitivities were quite different, with EC50 of 0.57 ng/mL for neonatal chondrocytes and EC50 of 8.70 ng/mL IGF-I for skeletally mature chondrocytes. These data suggest that IGF-I gene therapy may be more effective with younger cell sources. Both cell types were less sensitive to exogenous IGF-I than endogenous IGF-I.

  8. Differences in Cartilage-Forming Capacity of Expanded Human Chondrocytes From Ear and Nose and Their Gene Expression Profiles

    NARCIS (Netherlands)

    Hellingman, C.A.; Verwiel, E.T.P.; Slagt, I.; Koevoet, W.; Poublon, R.M.L.; Nolst-Trenite, G.J.; de Jong, R.J.B.; Jahr, H.; van Osch, G.J.V.M.

    2011-01-01

    The aim of this study was to evaluate the potential of culture-expanded human auricular and nasoseptal chondrocytes as cell source for regeneration of stable cartilage and to analyze the differences in gene expression profile of expanded chondrocytes from these specific locations. Auricular

  9. Differences in cartilage-forming capacity of expanded human chondrocytes from ear and nose and their gene expression profiles

    NARCIS (Netherlands)

    Hellingman, Catharine A.; Verwiel, Eugène T. P.; Slagt, Inez; Koevoet, Wendy; Poublon, René M. L.; Nolst-Trenité, Gilbert J.; Baatenburg de Jong, Robert J.; Jahr, Holger; van Osch, Gerjo J. V. M.

    2011-01-01

    The aim of this study was to evaluate the potential of culture-expanded human auricular and nasoseptal chondrocytes as cell source for regeneration of stable cartilage and to analyze the differences in gene expression profile of expanded chondrocytes from these specific locations. Auricular

  10. Overexpression of hsa-miR-148a promotes cartilage production and inhibits cartilage degradation by osteoarthritic chondrocytes

    NARCIS (Netherlands)

    Vonk, L A; Kragten, A H M; Dhert, W J A; Saris, D B F; Creemers, L B

    OBJECTIVE: Hsa-miR-148a expression is decreased in Osteoarthritis (OA) cartilage, but its functional role in cartilage has never been studied. Therefore, our aim was to investigate the effects of overexpressing hsa-miR-148a on cartilage metabolism of OA chondrocytes. DESIGN: OA chondrocytes were

  11. Overexpression of hsa-miR-148a promotes cartilage production and inhibits cartilage degradation by osteoarthritic chondrocytes

    NARCIS (Netherlands)

    Vonk, Lucienne A.; Kragten, Angela H.M.; Dhert, Wouter J.; Saris, Daniël B.F.; Creemers, Laura B.

    2014-01-01

    Objective Hsa-miR-148a expression is decreased in OA cartilage, but its functional role in cartilage has never been studied. Therefore, our aim was to investigate the effects of overexpressing hsa-miR-148a on cartilage metabolism of OA chondrocytes. Design OA chondrocytes were transfected with a

  12. Chondrocytes co-cultured with Stromal Vascular Fraction of adipose tissue present more intense chondrogenic characteristics than with Adipose Stem Cells

    NARCIS (Netherlands)

    Wu, Ling; Prins, H.J.; Leijten, Jeroen Christianus Hermanus; Helder, M.; Evseenko, D.; Moroni, L; van Blitterswijk, Clemens; Lin, Y.; Karperien, Hermanus Bernardus Johannes

    2016-01-01

    Partly replacement of chondrocytes by stem cells has been proposed to improve the performance of autologous chondrocytes implantation (ACI). Our previous studies showed that the increased cartilage production in pellet co-cultures of chondrocytes and mesenchymal stem cells (MSCs) is due to a trophic

  13. Andrographolide Enhances Proliferation and Prevents Dedifferentiation of Rabbit Articular Chondrocytes: An In Vitro Study

    Directory of Open Access Journals (Sweden)

    Li-ke Luo

    2015-01-01

    Full Text Available As the main active constituent of Andrographis paniculata that was applied in treatment of many diseases including inflammation in ancient China, andrographolide (ANDRO was found to facilitate reduction of edema and analgesia in arthritis. This suggested that ANDRO may be promising anti-inflammatory agent to relieve destruction and degeneration of cartilage after inflammation. In this study, the effect of ANDRO on rabbit articular chondrocytes in vitro was investigated. Results showed that not more than 8 μM ANDRO did no harm to chondrocytes (P0.05. The viability assay, hematoxylin-eosin, safranin O, and immunohistochemical staining also showed better performances in ANDRO groups. As to the doses, 3 μM ANDRO showed the best performance. The results indicate that ANDRO can accelerate proliferation of rabbit articular chondrocytes in vitro and meanwhile maintain the phenotype, which may provide valuable references for further exploration on arthritis.

  14. Stress relaxation analysis of single chondrocytes using porohyperelastic model based on AFM experiments

    Directory of Open Access Journals (Sweden)

    Trung Dung Nguyen

    2014-01-01

    Full Text Available Based on atomic force microscopytechnique, we found that the chondrocytes exhibits stress relaxation behavior. We explored the mechanism of this stress relaxation behavior and concluded that the intracellular fluid exuding out from the cells during deformation plays the most important role in the stress relaxation. We applied the inverse finite element analysis technique to determine necessary material parameters for porohyperelastic (PHE model to simulate stress relaxation behavior as this model is proven capable of capturing the non-linear behavior and the fluid-solid interaction during the stress relaxation of the single chondrocytes. It is observed that PHE model can precisely capture the stress relaxation behavior of single chondrocytes and would be a suitable model for cell biomechanics.

  15. ICAM-1 expression on chondrocytes in rheumatoid arthritis: induction by synovial cytokines

    Directory of Open Access Journals (Sweden)

    M. E. Davies

    1992-01-01

    Full Text Available The intercellular adhesion molecule-1 (ICAM-1 was found by immunostaining chondrocytes in cartilage from three patients with rheumatoid arthritis. Expression of ICAM-1 was restricted to chondrocytes in areas of erodedcartilage adjacent to the invading synovial tissue. Toluidine blue staining of these areas demonstrated severe depletion of the cartilage extracellular matrix. In areas of undamaged cartilage there was no ICAM-1 expression. Since ICAM-1 is not constitutively expressed on normal human articular cartilage, but could be induced in vitro by exogenous IL-1α, TNFα and IFNγ or by co-culturing cartilage with inflammatory rheumatoid synovium, we conclude that the induction of ICAM-1 on rheumatoid chondrocytes results from the synergistic action of a variety of cytokines produced by the inflammatory cells of the invading pannus.

  16. Fluoroquinolone's effect on growth of human chondrocytes and chondrosarcomas. In vitro and in vivo correlation

    DEFF Research Database (Denmark)

    Multhaupt, H A; Alvarez, J C; Rafferty, P A

    2001-01-01

    Clinical and in vitro studies have demonstrated that fluoroquinolones are toxic to chondrocytes; however, the exact mechanism of fluoroquinolone arthropathy is unknown. We investigated the toxicity of ciprofloxacin on normal cartilage and on cartilaginous tumors. Normal human cartilage, enchondroma...... with use of conventional light microscopy, electron microscopy, and immunohistochemistry to identify extracellular matrix, cell proliferation, and apoptosis. Cultures of normal chondrocytes expressed type-II collagen. Electron microscopy revealed a large amount of glycogen in the cells; the presence of fat...... of vimentin filaments. The treated chondrocytes showed a decrease in cell proliferation, but there was no induction of apoptosis or effect on the expression of extracellular matrix proteins. Ciprofloxacin-treated chondrosarcoma cultures and tissue samples showed changes in cartilage matrix composition...

  17. [REPORTING CRITICAL LAB RESULTS, A CHALLENGE FOR THE LAB AND THE PHYSICIAN - A SUMMARY OF FOUR YEARS OF EXPERIENCE IN MEIR MEDICAL CENTER LABORATORIES].

    Science.gov (United States)

    Rashid, Gloria; Goldman, Jacob; Weinstein, Doron; Tohami, Tali; Neumark, Eran; Weiss, Eli

    2015-08-01

    Critical laboratory results require prompt reporting to the attending physician, as they may indicate that a patient is in a life-threatening condition. Although this important subject has been covered in many publications, it needs more attention from our healthcare organizations, which have no official policy on the subject. Matching expectations between the doctor and the laboratory needs to be better defined. The aim of this work was to inform the community of doctors and laboratories about the multiple problems concerning the reporting of critical laboratory results, to create a platform for exchanging views and ideas, and to build an extensive infrastructure for developing a unified plan to address this important issue. We present the results of four years of experience of reporting critical laboratory values at the Meir Medical Center Laboratories. The idea leading this work was to present the relatively low rate of critical results reported by the laboratories in 2010, sharing the problems discovered while investigating the situation in depth, and presenting the solutions that enabled us to obtain the desired results within four years. Gradual implementation of these improvements resulted in critical value reporting increasing from 55% in 2010 to 95% currently. We suggest a model for improving critical laboratory values reporting based on our 4-year experience, which emphasizes: (1) The importance of selecting proper tests and values for critical results; (2) The significance of using technology and computerized measures to support the process; and (3) Developing quick procedures for monitoring and controlling the process.

  18. Grafting the alar rim: application as anatomical graft.

    Science.gov (United States)

    Gruber, Ronald P; Fox, Paige; Peled, Anne; Belek, Kyle A

    2014-12-01

    Alar rim contour and alar rim grafts have become essential components of rhinoplasty. Ideally, grafts of the nose should be anatomical in shape. So doing might make grafts of the alar rim more robust. The authors considered doing that by applying the graft as a continuous extension of the lateral crus. Twelve patients (two men and 10 women) constituted the study group (seven primary and five secondary cases). Of those, there were five concave rims, two concave rims with rim retraction, two boxy tips, and three cephalically oriented lateral crura. Surgical technique included the following: (1) an open approach was used; (2) a marginal incision that ignored the caudal margin of the lateral crus (the incision went straight posteriorly to a point 5 to 6 mm from the rim margin) was used; (3) a triangular graft was made to cover the exposed vestibular skin; (4) it was secured end to end to the caudal border of the lateral crus; and (5) the poster end was allowed to sit in a small subcutaneous pocket. Follow-up was 11 to 19 months. All 12 patients exhibited good rims as judged by a blinded panel. Rim retraction was not fully corrected in one patient, but no further treatment was required. One patient did require a secondary small rim graft for residual rim concavity. The concept of grafting the alar rim is strongly supported by the authors' results. The modifications the authors applied by designing the graft to be anatomical in shape has been a technical help.

  19. Autologous chondrocytes as a novel source for neo-chondrogenesis in haemophiliacs.

    Science.gov (United States)

    Stocco, Elena; Barbon, Silvia; Radossi, Paolo; Rajendran, Senthilkumar; Dalzoppo, Daniele; Bortolami, Marina; Bagno, Andrea; Grandi, Francesca; Gamba, Pier Giorgio; Parnigotto, Pier Paolo; Tagariello, Giuseppe; Grandi, Claudio

    2016-10-01

    Haemophilic arthropathy is the major cause of disability in patients with haemophilia and, despite prophylaxis with coagulation factor concentrates, some patients still develop articular complications. We evaluate the feasibility of a tissue engineering approach to improve current clinical strategies for cartilage regeneration in haemophiliacs by using autologous chondrocytes (haemophilic chondrocytes; HaeCs). Little is known about articular chondrocytes from haemophilic patients and no characterisation has as yet been performed. An investigation into whether blood exposure alters HaeCs should be interesting from the perspective of autologous implants. The typical morphology and expression of specific target genes and surface markers were therefore assessed by optical microscopy, reverse transcription plus the polymerase chain reaction (PCR), real-time PCR and flow-cytometry. We then considered chondrocyte behaviour on a bio-hybrid scaffold (based on polyvinyl alcohol/Wharton's jelly) as an in vitro model of articular cartilage prosthesis. Articular chondrocytes from non-haemophilic donors were used as controls. HaeC morphology and the resulting immunophenotype CD44(+)/CD49c(+)/CD49e(+)/CD151(+)/CD73(+)/CD49f(-)/CD26(-) resembled those of healthy donors. Moreover, HaeCs were active in the transcription of genes involved in the synthesis of the extracellular matrix proteins of the articular cartilage (ACAN, COL1A, COL2A, COL10A, COL9A, COMP, HAS1, SOX9), although the over-expression of COL1A1, COL10A1, COMP and HAS was observed. In parallel, the composite scaffold showed adequate mechanical and biological properties for cartilage tissue engineering, promoting chondrocyte proliferation. Our preliminary evidence contributes to the characterisation of HaeCs, highlighting the opportunity of using them for autologous cartilage implants in patients with haemophilia.

  20. Regulation of human mesenchymal stem cells differentiation into chondrocytes in extracellular matrix-based hydrogel scaffolds.

    Science.gov (United States)

    Du, Mingchun; Liang, Hui; Mou, Chenchen; Li, Xiaoran; Sun, Jie; Zhuang, Yan; Xiao, Zhifeng; Chen, Bing; Dai, Jianwu

    2014-02-01

    To induce human mesenchymal stem cells (hMSCs) to differentiate into chondrocytes in three-dimensional (3D) microenvironments, we developed porous hydrogel scaffolds using the cartilage extracellular matrix (ECM) components of chondroitin sulfate (CS) and collagen (COL). The turbidity and viscosity experiments indicated hydrogel could form through pH-triggered co-precipitation when pH=2-3. Enzyme-linked immunosorbent assay (ELISA) confirmed the hydrogel scaffolds could controllably release growth factors as envisaged. Transforming growth factor-β (TGF-β) was released to stimulate hMSCs differentiation into chondrocytes; and then collagen binding domain-basic fibroblast growth factor (CBD-bFGF) was released to improve the differentiation and preserve the chondrocyte phenotype. In in vitro cell culture experiments, the differentiation processes were compared in different microenvironments: 2D culture in culture plate as control, 3D culture in the fabricated scaffolds without growth factors (CC), the samples with CBD-bFGF (CC-C), the samples with TGF-β (CC-T), the samples with CBD-bFGF/TGF-β (CC-CT). Real-time polymerase chain reaction (RT-PCR) revealed the hMSC marker genes of CD44 and CD105 decreased; at the same time the chondrocyte marker genes of collagen type II and aggrecan increased, especially in the CC-CT sample. Immunostaining results further confirmed the hMSC marker protein of CD 44 disappeared and the chondrocyte marker protein of collagen type II emerged over time in the CC-CT sample. These results imply the ECM-based hydrogel scaffolds with growth factors can supply suitable 3D cell niches for hMSCs differentiation into chondrocytes and the differentiation process can be regulated by the controllably released growth factors. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. [Stimulation of maturing and terminal differentiation by concanavalin A in rabbit permanent chondrocyte cultures].

    Science.gov (United States)

    Yan, W Q; Yang, T S; Hou, L Z; Susuki, F; Kato, Y

    1994-12-01

    The effect of concanavalin A (Con A) on maturing and terminal differentiation in permanent chondrocyte cultures were examined. Chondrocytes isolated from permanent cartilage were seeded at low density and grown in MEM medium containing 10% fetal bovine serum, 50 micrograms/ml of ascorbic acid and antibiotics, at 37 degrees C under 50% CO2 in air. At 0.3% of low serum concentration, addition of Con A to the culture medium increased by 3- to 4-fold the incorporation of [35S] sulfate into large chondroitin sulfate proteoglycan that characteristically found in cartilage. Chemical analysis showed a 4-fold increase in the accumulation of macromolecular containing hexuronic acid in Con A-maintained cultures. The effect of Con A on [35S]sulfate incorporation into proteoglycan was greater than that of various growth factor or hormones. Brief exposure of the permanent chondrocytes to Con A (5 micrograms/ml) for 24 hours and subsequent incubation in its absence for 5-10 days resulted in 10- to 100-fold increase in alkaline phosphatase and binding of 1.25 (OH)2 vitamin D3 to cells. Treatment with Con A also resulted in 10- to 20-fold increase in calcium content and 45Ca incorporation into insoluble material. Methyl-D-mannopyranoside reversed the effect of Con A on [35S]sulfate incorporation into proteoglycan and alkaline phosphatase activity. Since other lectins, such as wheat germ agglutinin, lentil lectin, phytohemagglutinin, Ulex europeasu agglutinin and garden pea lectin had been tested to have little effect on [35S]sulfate incorporation into proteoglycans and induction of alkaline phosphatase activity, the Con A action on chondrocytes seems specific. These results indicate that Con A is a potent modulator of differentiation of chondrocytes, which induces the onset on a maturing and a terminal differentiation in chondrocytes, leading to extensive calcification of the extracellular matrix.

  2. High fat-diet and saturated fatty acid palmitate inhibits IGF-1 function in chondrocytes.

    Science.gov (United States)

    Nazli, S A; Loeser, R F; Chubinskaya, S; Willey, J S; Yammani, R R

    2017-09-01

    Insulin-like growth factor-1 (IGF-1) promotes matrix synthesis and cell survival in cartilage. Chondrocytes from aged and osteoarthritic cartilage have a reduced response to IGF-1. The purpose of this study was to determine the effect of free fatty acids (FFA) present in a high-fat diet on IGF-1 function in cartilage and the role of endoplasmic reticulum (ER) stress. C57BL/6 male mice were maintained on either a high-fat (60% kcal from fat) or a low-fat (10% kcal from fat) diet for 4 months. Mice were then sacrificed; femoral head cartilage caps were collected and treated with IGF-1 to measure proteoglycan (PG) synthesis. Cultured human chondrocytes were treated with 500 μM FFA palmitate or oleate, followed by stimulation with (100 ng/ml) IGF-1 overnight to measure CHOP (a protein marker for ER stress) and PG synthesis. Human chondrocytes were pre-treated with palmitate or 1 mM 4-phenyl butyric acid (PBA) or 1 μM C-Jun N terminal Kinase (JNK) inhibitor, and IGF-1 function (PG synthesis and signaling) was measured. Cartilage explants from mice on the high fat-diet showed reduced IGF-1 mediated PG synthesis compared to a low-fat group. Treatment of human chondrocytes with palmitate induced expression of CHOP, activated JNK and inhibited IGF-1 function. PBA, a small molecule chemical chaperone that alleviates ER stress rescued IGF-1 function and a JNK inhibitor rescued IGF-1 signaling. Palmitate-induced ER stress inhibited IGF-1 function in chondrocytes/cartilage via activating the mitogen-activated protein (MAP) kinase JNK. This is the first study to demonstrate that ER stress is metabolic factor that regulates IGF-1 function in chondrocytes. Copyright © 2017 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

  3. Curcumin Inhibits Apoptosis of Chondrocytes through Activation ERK1/2 Signaling Pathways Induced Autophagy

    Directory of Open Access Journals (Sweden)

    Xiaodong Li

    2017-04-01

    Full Text Available Osteoarthritis (OA is an inflammatory disease of load-bearing synovial joints that is currently treated with drugs that exhibit numerous side effects and are only temporarily effective in treating pain, the main symptom of the disease. Consequently, there is an acute need for novel, safe, and more effective chemotherapeutic agents for the treatment of osteoarthritis and related arthritic diseases. Curcumin, the principal curcuminoid and the most active component in turmeric, is a biologically active phytochemical. Evidence from several recent in vitro studies suggests that curcumin may exert a chondroprotective effect through actions such as anti-inflammatory, anti-oxidative stress, and anti-catabolic activity that are critical for mitigating OA disease pathogenesis and symptoms. In the present study, we investigated the protective mechanisms of curcumin on interleukin 1β (IL-1β-stimulated primary chondrocytes in vitro. The treatment of interleukin (IL-1β significantly reduces the cell viability of chondrocytes in dose and time dependent manners. Co-treatment of curcumin with IL-1β significantly decreased the growth inhibition. We observed that curcumin inhibited IL-1β-induced apoptosis and caspase-3 activation in chondrocytes. Curcumin can increase the expression of phosphorylated extracellular signal-regulated kinases 1/2 (ERK1/2, autophagy marker light chain 3 (LC3-II, and Beclin-1 in chondrocytes. The expression of autophagy markers could be decreased when the chondrocytes were incubated with ERK1/2 inhibitor U0126. Our results suggest that curcumin suppresses apoptosis and inflammatory signaling through its actions on the ERK1/2-induced autophagy in chondrocytes. We propose that curcumin should be explored further for the prophylactic treatment of osteoarthritis in humans and companion animals.

  4. Suction blister grafting - Modifications for easy harvesting and grafting

    Directory of Open Access Journals (Sweden)

    2012-01-01

    Full Text Available Suction blister grafting is a simple modality of treatment of patients with resistant and stable vitiligo. But raising the blisters may be time consuming and transferring to the recipient site may be difficult as the graft is ultrathin. By doing some modifications we can make the technique simpler and easier. We can decrease the blister induction time by intradermal injection of saline, exposure to Wood′s lamp, intrablister injection of saline. By these methods we can decrease the blister induction time from 2-3 hrs to 45-90 minutes. After harvesting the graft, it can be transferred to the recipient area by taking the graft on a sterile glass slide, on the gloved finger, rolling the graft over a sterile syringe and then spreading on the recipient area, or taking on the sterile wrapper of paraffin dressing and then placing over the recipient area.

  5. The Results of Fetal Chondrocytes Transplantation in Patients with Rheumatoid Arthritis

    Directory of Open Access Journals (Sweden)

    Natalya Krivoruchko

    2014-12-01

    Full Text Available Introduction. Nowadays anti-inflammatory and immunosuppressive therapy has significantly improved the quality of life and prognosis of rheumatoid arthritis (RA. Nevertheless, there are still many patients with progressive rheumatoid inflammation, resulting in the destruction of joints. Cell therapy seems like a promising direction in rheumatology. The aim of our research was to evaluate the efficacy of fetal chondrocyte transplantation in patients with RA.Methods. We examined 60 patients with rheumatoid arthritis (I - III stages between 20 and 63 years of age. They were divided into 2 groups: the first group underwent the fetal chondrocytes transplantation (n = 40, and the second was a control group who got conservative therapy (n = 20. Donor cells were taken from the chondrogenic layer of the humerus or femur heads and hip condyles of human embryos in gestation for 17-20 weeks. A suspension of fetal chondrocytes injected into affected areas of the articular surfaces under X-ray control. Cell viability was determined before the injection. Efficacy of the therapy was assessed by clinical, instrumental, and laboratory tests. This clinical trial was allowed by The Ministry of Public Health and Ethics Committee. All of our patients gave informed consent for the fetal chondrocytes transplantation.Results. Evaluation of the clinical manifestations of RA in the first group of patients showed 3.7 times decrease in pain and 1.6 times relief of synovitis. Complete reduction of contracture was observed in 82% of patients in the first group. Morphometric changes in X-ray demonstrated inhibition of the destruction in articular cartilage and surfaces of bones after transplantation of fetal chondrocytes. The dynamics of morphological changes in synovium showed 2.5 times reduction of the inflammatory reaction. Transplantation of fetal chondrocytes led to a significant reduction in ESR, CRP, fibrinogen , γ-globulin after a period of 12 months (p < 0

  6. CCAAT/Enhancer Binding Protein β Regulates Expression of Indian Hedgehog during Chondrocytes Differentiation

    Science.gov (United States)

    Ushijima, Takahiro; Okazaki, Ken; Tsushima, Hidetoshi; Ishihara, Kohei; Doi, Toshio; Iwamoto, Yukihide

    2014-01-01

    Background CCAAT/enhancer binding protein β (C/EBPβ) is a transcription factor that promotes hypertrophic differentiation of chondrocytes. Indian hedgehog (Ihh) also stimulates the hypertrophic transition of chondrocytes. Furthermore, runt-related transcription factor-2 (RUNX2) was reported to regulate chondrocyte maturation during skeletal development and to directly regulate transcriptional activity of Ihh. In this study, we investigated whether the interaction of C/EBPβ and RUNX2 regulates the expression of Ihh during chondrocyte differentiation. Methodology/Results Immunohistochemistry of embryonic growth plate revealed that both C/EBPβ and Ihh were strongly expressed in pre-hypertrophic and hypertrophic chondrocytes. Overexpression of C/EBPβ by adenovirus vector in ATDC5 cells caused marked stimulation of Ihh and Runx2. Conversely, knockdown of C/EBPβ by lentivirus expressing shRNA significantly repressed Ihh and Runx2 in ATDC5 cells. A reporter assay revealed that C/EBPβ stimulated transcriptional activity of Ihh. Deletion and mutation analysis showed that the C/EBPβ responsive element was located between −214 and −210 bp in the Ihh promoter. An electrophoretic mobility shift assay (EMSA) and a chromatin immunoprecipitation (ChIP) assay also revealed the direct binding of C/EBPβ to this region. Moreover, reporter assays demonstrated that RUNX2 failed to stimulate the transcriptional activity of the Ihh promoter harboring a mutation at the C/EBPβ binding site. EMSA and ChIP assays showed that RUNX2 interacted to this element with C/EBPβ. Immunoprecipitation revealed that RUNX2 and C/EBPβ formed heterodimer complex with each other in the nuclei of chondrocytes. These data suggested that the C/EBPβ binding element is also important for RUNX2 to regulate the expression of Ihh. Ex vivo organ culture of mouse limbs transfected with C/EBPβ showed that the expression of Ihh and RUNX2 was increased upon ectopic C/EBPβ expression. Conclusions C

  7. CCAAT/enhancer binding protein β regulates expression of Indian hedgehog during chondrocytes differentiation.

    Directory of Open Access Journals (Sweden)

    Takahiro Ushijima

    Full Text Available CCAAT/enhancer binding protein β (C/EBPβ is a transcription factor that promotes hypertrophic differentiation of chondrocytes. Indian hedgehog (Ihh also stimulates the hypertrophic transition of chondrocytes. Furthermore, runt-related transcription factor-2 (RUNX2 was reported to regulate chondrocyte maturation during skeletal development and to directly regulate transcriptional activity of Ihh. In this study, we investigated whether the interaction of C/EBPβ and RUNX2 regulates the expression of Ihh during chondrocyte differentiation.Immunohistochemistry of embryonic growth plate revealed that both C/EBPβ and Ihh were strongly expressed in pre-hypertrophic and hypertrophic chondrocytes. Overexpression of C/EBPβ by adenovirus vector in ATDC5 cells caused marked stimulation of Ihh and Runx2. Conversely, knockdown of C/EBPβ by lentivirus expressing shRNA significantly repressed Ihh and Runx2 in ATDC5 cells. A reporter assay revealed that C/EBPβ stimulated transcriptional activity of Ihh. Deletion and mutation analysis showed that the C/EBPβ responsive element was located between -214 and -210 bp in the Ihh promoter. An electrophoretic mobility shift assay (EMSA and a chromatin immunoprecipitation (ChIP assay also revealed the direct binding of C/EBPβ to this region. Moreover, reporter assays demonstrated that RUNX2 failed to stimulate the transcriptional activity of the Ihh promoter harboring a mutation at the C/EBPβ binding site. EMSA and ChIP assays showed that RUNX2 interacted to this element with C/EBPβ. Immunoprecipitation revealed that RUNX2 and C/EBPβ formed heterodimer complex with each other in the nuclei of chondrocytes. These data suggested that the C/EBPβ binding element is also important for RUNX2 to regulate the expression of Ihh. Ex vivo organ culture of mouse limbs transfected with C/EBPβ showed that the expression of Ihh and RUNX2 was increased upon ectopic C/EBPβ expression.C/EBPβ and RUNX2 cooperatively stimulate

  8. Gli3 acts as a repressor downstream of Ihh in regulating two distinct steps of chondrocyte differentiation.

    Science.gov (United States)

    Koziel, Lydia; Wuelling, Manuela; Schneider, Sabine; Vortkamp, Andrea

    2005-12-01

    During endochondral ossification, the secreted growth factor Indian hedgehog (Ihh) regulates several differentiation steps. It interacts with a second secreted factor, parathyroid hormone-related protein (PTHrP), to regulate the onset of hypertrophic differentiation, and it regulates chondrocyte proliferation and ossification of the perichondrium independently of PTHrP. To investigate how the Ihh signal is translated in the different target tissues, we analyzed the role of the zinc-finger transcription factor Gli3, which acts downstream of hedgehog signals in other organs. Loss of Gli3 in Ihh mutants restores chondrocyte proliferation and delays the accelerated onset of hypertrophic differentiation observed in Ihh-/- mutants. Furthermore the expression of the Ihh target genes patched (Ptch) and PTHrP is reactivated in Ihh-/-;Gli3-/- mutants. Gli3 seems thus to act as a strong repressor of Ihh signals in regulating chondrocyte differentiation. In addition, loss of Gli3 in mice that overexpress Ihh in chondrocytes accelerates the onset of hypertrophic differentiation by reducing the domain and possibly the level of PTHrP expression. Careful analysis of chondrocyte differentiation in Gli3-/- mutants revealed that Gli3 negatively regulates the differentiation of distal, low proliferating chondrocytes into columnar, high proliferating cells. Our results suggest a model in which the Ihh/Gli3 system regulates two distinct steps of chondrocyte differentiation: (1) the switch from distal into columnar chondrocytes is repressed by Gli3 in a PTHrP-independent mechanism; (2) the transition from proliferating into hypertrophic chondrocytes is regulated by Gli3-dependent expression of PTHrP. Furthermore, by regulating distal chondrocyte differentiation, Gli3 seems to position the domain of PTHrP expression.

  9. Impact of human platelet lysate on the expansion and chondrogenic capacity of cultured human chondrocytes for cartilage cell therapy.

    Science.gov (United States)

    Sykes, J G; Kuiper, J H; Richardson, J B; Roberts, S; Wright, K T; Kuiper, N J

    2018-05-01

    High hopes have been pinned on regenerative medicine strategies in order to prevent the progression of cartilage damage to osteoarthritis, particularly by autologous chondrocyte implantation (ACI). The loss of chondrocyte phenotype during in vitro monolayer expansion, a necessary step to obtain sufficient cell numbers, may be a key limitation in ACI. In this study, it was determined whether a shorter monolayer expansion approach could improve chondrogenic differentiation. The effects of two supplement types, foetal bovine serum (FBS) and Stemulate™ (a commercial source of human platelet lysate), on the expansion and re-differentiation potential of human chondrocytes, isolated from five individuals, were compared. Chondrocytes were expanded with 10 % FBS or 10 % Stemulate™. Pellets were cultured for 28 d in chondrogenic differentiation medium and assessed for the presence of cartilage matrix molecules and genes associated with chondrogenicity. Stemulate™ significantly enhanced the proliferation rate [average population doubling times: FBS, 25.07 ± 6.98 d (standard error of the mean, SEM) vs. Stemulate™, 13.10 ± 2.57 d (SEM)]. Sulphated glycosaminoglycans (sGAG), total collagen and qRT-PCR analyses of cartilage genes showed that FBS-expanded chondrocytes demonstrated significantly better chondrogenic capacity than Stemulate™-expanded chondrocytes. Histologically, FBS-expanded chondrocyte pellets appeared to be more stable, with a more intense staining for toluidine blue, indicating a greater chondrogenic capacity. Although Stemulate™ positively influenced chondrocyte proliferation, it had a negative effect on chondrogenic differentiation potential. This suggested that, in the treatment of cartilage defects, Stemulate™ might not be the ideal supplement for expanding chondrocytes (which maintained a chondrocyte phenotype) and, hence, for cell therapies (including ACI).

  10. Impact of human platelet lysate on the expansion and chondrogenic capacity of cultured human chondrocytes for cartilage cell therapy

    Directory of Open Access Journals (Sweden)

    JG Sykes

    2018-05-01

    Full Text Available High hopes have been pinned on regenerative medicine strategies in order to prevent the progression of cartilage damage to osteoarthritis, particularly by autologous chondrocyte implantation (ACI. The loss of chondrocyte phenotype during in vitro monolayer expansion, a necessary step to obtain sufficient cell numbers, may be a key limitation in ACI. In this study, it was determined whether a shorter monolayer expansion approach could improve chondrogenic differentiation. The effects of two supplement types, foetal bovine serum (FBS and Stemulate™ (a commercial source of human platelet lysate, on the expansion and re-differentiation potential of human chondrocytes, isolated from five individuals, were compared. Chondrocytes were expanded with 10 % FBS or 10 % Stemulate™. Pellets were cultured for 28 d in chondrogenic differentiation medium and assessed for the presence of cartilage matrix molecules and genes associated with chondrogenicity. Stemulate™ significantly enhanced the proliferation rate [average population doubling times: FBS, 25.07 ± 6.98 d (standard error of the mean, SEM vs. Stemulate™, 13.10 ± 2.57 d (SEM]. Sulphated glycosaminoglycans (sGAG, total collagen and qRT-PCR analyses of cartilage genes showed that FBS-expanded chondrocytes demonstrated significantly better chondrogenic capacity than Stemulate™-expanded chondrocytes. Histologically, FBS-expanded chondrocyte pellets appeared to be more stable, with a more intense staining for toluidine blue, indicating a greater chondrogenic capacity. Although Stemulate™ positively influenced chondrocyte proliferation, it had a negative effect on chondrogenic differentiation potential. This suggested that, in the treatment of cartilage defects, Stemulate™ might not be the ideal supplement for expanding chondrocytes (which maintained a chondrocyte phenotype and, hence, for cell therapies (including ACI.

  11. SOME TECHNIQUES IN CORNEAL GRAFTING

    African Journals Online (AJOL)

    1971-04-10

    Apr 10, 1971 ... current herpes corneae. The visual acuity was less than. 6/60. The left eye had had a central nebula since child- hood and was deemed amblyopic. Six weeks after a 7 x 0·3 mm lamellar graft in the right eye was placed, ulceration occurred in the graft junction. A total thin conjunctival flap was sutured over.

  12. Polyether/Polyester Graft Copolymers

    Science.gov (United States)

    Bell, Vernon L., Jr.; Wakelyn, N.; Stoakley, D. M.; Proctor, K. M.

    1986-01-01

    Higher solvent resistance achieved along with lower melting temperature. New technique provides method of preparing copolymers with polypivalolactone segments grafted onto poly (2,6-dimethyl-phenylene oxide) backbone. Process makes strong materials with improved solvent resistance and crystalline, thermally-reversible crosslinks. Resulting graft copolymers easier to fabricate into useful articles, including thin films, sheets, fibers, foams, laminates, and moldings.

  13. Radiation grafting on natural films

    Science.gov (United States)

    Lacroix, M.; Khan, R.; Senna, M.; Sharmin, N.; Salmieri, S.; Safrany, A.

    2014-01-01

    Different methods of polymer grafting using gamma irradiation are reported in the present study for the preparation of newly functionalized biodegradable films, and some important properties related to their mechanical and barrier properties are described. Biodegradable films composed of zein and poly(vinyl alcohol) (PVA) were gamma-irradiated in presence of different ratios of acrylic acid (AAc) monomer for compatibilization purpose. Resulting grafted films (zein/PVA-g-AAc) had their puncture strength (PS=37-40 N mm-1) and puncture deformation (PD=6.5-9.8 mm) improved for 30% and 50% PVA in blend, with 5% AAc under 20 kGy. Methylcellulose (MC)-based films were irradiated in the presence of 2-hydroxyethyl methacrylate (HEMA) or silane, in order to determine the effect of monomer grafting on the mechanical properties of films. It was found that grafted films (MC-g-HEMA and MC-g-silane) using 35% monomer performed higher mechanical properties with PS values of 282-296 N mm-1 and PD of 5.0-5.5 mm under 10 kGy. Compatibilized polycaprolactone (PCL)/chitosan composites were developed via grafting silane in chitosan films. Resulting trilayer grafted composite film (PCL/chitosan-g-silane/PCL) presented superior tensile strength (TS=22 MPa) via possible improvement of interfacial adhesion (PCL/chitosan) when using 25% silane under 10 kGy. Finally, MC-based films containing crystalline nanocellulose (CNC) as a filling agent were prepared and irradiated in presence of trimethylolpropane trimethacrylate (TMPTMA) as a grafted plasticizer. Grafted films (MC-g-TMPTMA) presented superior mechanical properties with a TS of 47.9 MPa and a tensile modulus (TM) of 1792 MPa, possibly due to high yield formation of radicals to promote TMPTMA grafting during irradiation. The addition of CNC led to an additional improvement of the barrier properties, with a significant 25% reduction of water vapor permeability (WVP) of grafted films.

  14. Correlation Between Clinical and Radiological Outcomes After Matrix-Induced Autologous Chondrocyte Implantation in the Femoral Condyles.

    Science.gov (United States)

    Ebert, Jay R; Smith, Anne; Fallon, Michael; Wood, David J; Ackland, Timothy R

    2014-08-01

    Matrix-induced autologous chondrocyte implantation (MACI) is an established technique for the repair of knee chondral defects, although the correlation between clinical and radiological outcomes after surgery is poorly understood. To determine the correlation between clinical and radiological outcomes throughout the postoperative timeline to 5 years after MACI. Cohort study (diagnosis); Level of evidence, 3. This retrospective study was undertaken in 83 patients (53 male, 30 female) with complete clinical and radiological follow-up at 1, 2, and 5 years after MACI. The mean age of patients was 38.9 years (range, 13-62 years), with a mean body mass index (BMI) of 26.6 kg/m(2) (range, 16.8-34.8 kg/m(2)), mean defect size of 3.3 cm(2) (range, 1-9 cm(2)), and mean preoperative duration of symptoms of 9.2 years (range, 1-46 years). Patients indicated for MACI in this follow-up were 13 to 65 years of age, although they were excluded if they had a BMI >35 kg/m(2), had undergone prior extensive meniscectomy, or had ongoing progressive inflammatory arthritis. Patients were assessed clinically using the Knee Injury and Osteoarthritis Outcome Score (KOOS). Magnetic resonance imaging (MRI) was used to evaluate the graft using a 1.5-T or 3-T clinical scanner; the MRI assessment included 8 parameters of graft repair (infill, signal intensity, border integration, surface contour, structure, subchondral lamina, subchondral bone, and effusion) based on the magnetic resonance observation of cartilage repair tissue (MOCART) score as well as an MRI composite score. The degree of an association between the MRI parameters and the KOOS subscales at each postoperative time point was assessed with the Spearman correlation coefficient (SCC), and significance was determined at P correlations over time and statistically significant associations at 5 years with KOOS-Pain (SCC, 0.25; P = .020), KOOS-Activities of Daily Living (SCC, 0.26; P = .018), and KOOS-Sport (SCC, 0.32; P = .003). Apart

  15. TNF/TNFR1 pathway and endoplasmic reticulum stress are involved in ofloxacin-induced apoptosis of juvenile canine chondrocytes

    International Nuclear Information System (INIS)

    Zhang, Fu-Tao; Ding, Yi; Shah, Zahir; Xing, Dan; Gao, Yuan; Liu, Dong Ming; Ding, Ming-Xing

    2014-01-01

    Background and purpose: Quinolones cause obvious cartilaginous lesions in juvenile animals by chondrocyte apoptosis, which results in the restriction of their use in pediatric and adolescent patients. Studies showed that chondrocytes can be induced to produce TNFα, and the cisternae of the endoplasmic reticulum in quinolone-treated chondrocytes become dilated. We investigated whether TNF/TNFR 1 pathway and endoplasmic reticulum stress (ERs) are involved in ofloxacin (a typical quinolone)-induced apoptosis of juvenile canine chondrocytes. Experimental approach: Canine juvenile chondrocytes were treated with ofloxacin. Cell survival and apoptosis rates were determined with MTT method and flow cytometry, respectively. The gene expression levels of the related signaling molecules (TNFα, TNFR 1 , TRADD, FADD and caspase-8) in death receptor pathways and main apoptosis-related molecules (calpain, caspase-12, GADD153 and GRP78) in ERs were measured by qRT-PCR. The gene expression of TNFR 1 was suppressed with its siRNA. The protein levels of TNFα, TNFR 1 and caspase-12 were assayed using Western blotting. Key results: The survival rates decreased while apoptosis rates increased after the chondrocytes were treated with ofloxacin. The mRNA levels of the measured apoptosis-related molecules in death receptor pathways and ERs, and the protein levels of TNFα, TNFR 1 and caspase-12 increased after the chondrocytes were exposed to ofloxacin. The downregulated mRNA expressions of TNFR 1 , Caspase-8 and TRADD, and the decreased apoptosis rates of the ofloxacin-treated chondrocytes occurred after TNFR 1 –siRNA interference. Conclusions and implications: Ofloxacin-induced chondrocyte apoptosis in a time- and concentration-dependent fashion. TNF/TNFR 1 pathway and ERs are involved in ofloxacin-induced apoptosis of juvenile canine chondrocytes in the early stage. - Highlights: • Chondrocyte apoptosis is induced by ofloxacin in a time- and concentration-dependent manners.

  16. Impact of routine rotavirus vaccination on all-cause and rotavirus hospitalizations during the first four years following vaccine introduction in Rwanda.

    Science.gov (United States)

    Sibomana, Hassan; Rugambwa, Celse; Mwenda, Jason M; Sayinzoga, Felix; Iraguha, Gisele; Uwimana, Jeanine; Parashar, Umesh D; Tate, Jacqueline E

    2018-05-10

    Rwanda introduced pentavalent rotavirus vaccine into its national immunization program in 2012. To determine the long-term impact of rotavirus vaccine on disease burden in a high burden setting, we examined trends in rotavirus and all-cause diarrhea hospitalizations in the first four years following rotavirus vaccine introduction. We used data from an active surveillance system, from a review of pediatric ward registries, and from the Health Management Information System to describe trends in rotavirus and all-cause diarrhea hospitalizations from January 2009 through December 2016. Percent reductions were calculated to compare the number of all-cause and rotavirus diarrhea hospitalizations pre- and post-rotavirus vaccine introduction. The proportion of diarrhea hospitalizations due to rotavirus declined by 25-44% among all children introduction to 12-13% post-vaccine introduction. In the national hospital discharge data, substantial decreases were observed in all-cause diarrhea hospitalizations among children introduction era. Published by Elsevier Ltd.

  17. Central odontogenic fibroma (simple type) in a four year old boy: Atypical cone-beam computed tomographic appearance with periosteal reaction

    Energy Technology Data Exchange (ETDEWEB)

    Anbiaee, Najme; Ebrahimnejad, Hamed; Sanaei, Alireza [Dept. of , Oral and Maxillofacial Radiology, Maxillofacial Diseases Research Center, School of Dentistry, Mashhad University of Medical Sciences, Mashhad (Iran, Islamic Republic of)

    2015-06-15

    Central odontogenic fibroma (COF) is a rare benign tumor that accounts for 0.1% of all odontogenic tumors. A case of COF (simple type) of the mandible in a four-year-old boy is described in this report. The patient showed asymptomatic swelling in the right inferior border of the lower jaw for one week. A panoramic radiograph showed a poorly-defined destructive unilocular radiolucent area. Cone-beam computed tomography showed expansion and perforation of the adjacent cortical bone plates. A periosteal reaction with the Codman triangle pattern was clearly visible in the buccal cortex. Since the tumor had destroyed a considerable amount of bone, surgical resection was performed. No recurrence was noted.

  18. Comprehensive Survey Results of Childhood Thyroid Ultrasound Examinations in Fukushima in the First Four Years After the Fukushima Daiichi Nuclear Power Plant Accident.

    Science.gov (United States)

    Suzuki, Shinichi; Suzuki, Satoru; Fukushima, Toshihiko; Midorikawa, Sanae; Shimura, Hiroki; Matsuzuka, Takashi; Ishikawa, Tetsuo; Takahashi, Hideto; Ohtsuru, Akira; Sakai, Akira; Hosoya, Mitsuaki; Yasumura, Seiji; Nollet, Kenneth E; Ohira, Tetsuya; Ohto, Hitoshi; Abe, Masafumi; Kamiya, Kenji; Yamashita, Shunichi

    2016-06-01

    Thyroid nodules and cancers are rare in children compared with adults. However, after the 1986 Chernobyl Nuclear Power Plant accident, a rapid increase in childhood thyroid cancer was observed. To avoid any confusion and misunderstanding of data obtained in Fukushima after the 2011 nuclear accident, baseline prevalence of thyroid nodules and cancers should be carefully assessed with standardized criteria systematically, and comprehensively applied to the population perceived to be at risk. Under the official framework of the Fukushima Health Management Survey, the thyroids of children in Fukushima were examined using ultrasound, and the results collected in the first four years after the nuclear accident were analyzed in order to establish a baseline prevalence of childhood thyroid abnormalities, especially cancer. Of 367,685 people aged 18 years or younger as of April 1, 2011, who were living in Fukushima Prefecture at the time of the accident, 300,476 underwent thyroid ultrasound screening. Of those, 2108 subjects with thyroid nodules were further examined using an advanced ultrasound instrument, with standardized criteria applied to determine the need for fine-needle aspiration cytology (FNAC). FNAC results determined the need for surgery and histological confirmation of the cytological diagnosis. Of the 2108 rescreened subjects, 543 underwent FNAC, of whom 113 were diagnosed with malignancy or suspected malignancy. Subsequently, 99 patients underwent surgical resection, revealing 95 cases of papillary thyroid cancer, three poorly differentiated cancers, and one benign nodule. The overall prevalence of childhood thyroid cancer in Fukushima was determined to be 37.3 per 100,000 with no significant differences between evacuated and non-evacuated areas. Thyroid cancer patients had external exposure estimates of detected in this four-year study in Fukushima can be attributed to mass screening. It clearly exceeds what is found incidentally anywhere else. Direct

  19. Influence of bone morphogenetic protein-2 on the extracellular matrix, material properties, and gene expression of long-term articular chondrocyte cultures: loss of chondrocyte stability.

    Science.gov (United States)

    Krawczak, David A; Westendorf, Jennifer J; Carlson, Cathy S; Lewis, Jack L

    2009-06-01

    The aim of this study was to determine the effects of bone morphogenetic protein-2 (BMP-2) on articular chondrocyte tissues grown as monolayers in vitro for up to 8 weeks. Articular chondrocytes were isolated from New Zealand White rabbits and plated in monolayer cultures. The cultures were supplemented with 100 ng/mL of BMP-2 for up to 8 weeks and the extracellular matrix (ECM) composition, material properties, and messenger RNA (mRNA) expression were analyzed. mRNA expression of cartilage-specific genes, type II collagen, and aggrecan showed that BMP-2 enhanced chondrocyte stability for up to 3 weeks. After 3 weeks in culture, there was substantially more type I collagen expression and more osteopontin and runt-related transcription factor 2 expression in 5- and 8-week cultures treated with BMP-2 than in controls. Additionally, matrix metalloproteinase-13 and ADAMTS-5 (A disintegrin-like and metalloproteinase with thrombospondin 5) were upregulated in 5- and 8-week cultures treated with BMP-2, coinciding with a loss of ECM density, collagen, and proteoglycan. Eight-week tissue stimulated with BMP-2 was more fragile and tore more easily when removed from the culture dish as compared to controls, suggesting temporal limitations to the effectiveness of BMP-2 in monolayer systems and perhaps other models to enhance the generation of a cartilage-like tissue for tissue engineering purposes.

  20. Photodynamic damage to cartilage and synovial tissue grafted on a chick's chorioallantoic membrane

    Science.gov (United States)

    Fisher, M.; Nahir, A. M.; Kimel, Sol

    1997-09-01

    Rheumatoid arthritis (RA) is a chronic inflammatory disease of the synovial joints causing pain deformities and disability. The highly vascular inflamed synovium has aggressive and destructive characteristics, it invades, erodes and gradually destroys cartilage and underlying bone. Photodynamic therapy (PDT) was performed using the chick chorioallantoic membrane (CAM) model to investigate the vitality of synovium and cartilage implanted on the CAM. Synovium, obtained from human patients, was grafted onto the CAM; gross microscopy and histology proved its vitality 7 days post grafting. Cartilage obtained from rabbit knee joint was also maintained on the CAM for 7 days. Its vitality was demonstrated by histology and by measuring metabolic and enzymatic activity of cartilage cells (chondrocytes) as well as the collagen and proteoglycans content. Selective PDT was performed using aluminum phthalocyanine tetrasulfonate (AlPcS4), a hydrophilic compound, soluble in biological solutions, as a photosensitizer. After irradiation with a diode laser (lambda equals 670 nm, 10 mW) damage was observed in vascularized synovium grafts, whereas avascular cartilage remained intact.

  1. Graft intolerance syndrome requiring graft nephrectomy after late kidney graft failure: can it be predicted? A retrospective cohort study.

    Science.gov (United States)

    Bunthof, Kim L W; Verhoeks, Carmen M; van den Brand, Jan A J G; Hilbrands, Luuk B

    2018-02-01

    Graft nephrectomy is recommended in case of early graft failure. When the graft fails more than 3-6 months after transplantation, it is current practice to follow a wait-and-see policy. A common indication for graft removal is the graft intolerance syndrome. We aimed to create a risk prediction model for the occurrence of graft intolerance resulting in graft nephrectomy. We collected data of kidney transplantations performed in our center between 1980 and 2010 that failed at least 6 months after transplantation. We evaluated the association between baseline characteristics and the occurrence of graft nephrectomy because of graft intolerance using a competing risk regression model. Prognostic factors were included in a multivariate prediction model. In- and exclusion criteria were met in 288 cases. In 48 patients, the graft was removed because of graft intolerance. Donor age, the number of rejections, and shorter graft survival were predictive factors for graft nephrectomy because of the graft intolerance syndrome. These factors were included in a prediction rule. Using donor age, graft survival, and the number of rejections, clinicians can predict the need for graft nephrectomy with a reasonable accuracy. © 2017 Steunstichting ESOT.

  2. Surviving endoplasmic reticulum stress is coupled to altered chondrocyte differentiation and function.

    Directory of Open Access Journals (Sweden)

    Kwok Yeung Tsang

    2007-03-01

    Full Text Available In protein folding and secretion disorders, activation of endoplasmic reticulum (ER stress signaling (ERSS protects cells, alleviating stress that would otherwise trigger apoptosis. Whether the stress-surviving cells resume normal function is not known. We studied the in vivo impact of ER stress in terminally differentiating hypertrophic chondrocytes (HCs during endochondral bone formation. In transgenic mice expressing mutant collagen X as a consequence of a 13-base pair deletion in Col10a1 (13del, misfolded alpha1(X chains accumulate in HCs and elicit ERSS. Histological and gene expression analyses showed that these chondrocytes survived ER stress, but terminal differentiation is interrupted, and endochondral bone formation is delayed, producing a chondrodysplasia phenotype. This altered differentiation involves cell-cycle re-entry, the re-expression of genes characteristic of a prehypertrophic-like state, and is cell-autonomous. Concomitantly, expression of Col10a1 and 13del mRNAs are reduced, and ER stress is alleviated. ERSS, abnormal chondrocyte differentiation, and altered growth plate architecture also occur in mice expressing mutant collagen II and aggrecan. Alteration of the differentiation program in chondrocytes expressing unfolded or misfolded proteins may be part of an adaptive response that facilitates survival and recovery from the ensuing ER stress. However, the altered differentiation disrupts the highly coordinated events of endochondral ossification culminating in chondrodysplasia.

  3. Inhibition of cyclooxygenase-2 impacts chondrocyte hypertrophic differentiation during endochondral ossification

    Directory of Open Access Journals (Sweden)

    TJM Welting

    2011-12-01

    Full Text Available Skeletogenesis and bone fracture healing involve endochondral ossification, a process during which cartilaginous primordia are gradually replaced by bone tissue. In line with a role for cyclooxygenase-2 (COX-2 in the endochondral ossification process, non-steroidal anti-inflammatory drugs (NSAIDs were reported to negatively affect bone fracture healing due to impaired osteogenesis. However, a role for COX-2 activity in the chondrogenic phase of endochondral ossification has not been addressed before. We show that COX-2 activity fulfils an important regulatory function in chondrocyte hypertrophic differentiation. Our data reveal essential cross-talk between COX-2 and bone morphogenic protein-2 (BMP-2 during chondrocyte hypertrophic differentiation. BMP-2 mediated chondrocyte hypertrophy is associated with increased COX-2 expression and pharmacological inhibition of COX-2 activity by NSAIDs (e.g., Celecoxib decreases hypertrophic differentiation in various chondrogenic models in vitro and in vivo, while leaving early chondrogenic development unaltered. Our findings demonstrate that COX-2 activity is a novel factor partaking in chondrocyte hypertrophy in the context of endochondral ossification and these observations provide a novel etiological perspective on the adverse effects of NSAIDs on bone fracture healing and have important implications for the use of NSAIDs during endochondral skeletal development.

  4. Effects of Bauhinia championii (Benth.) Benth. polysaccharides on the proliferation and cell cycle of chondrocytes.

    Science.gov (United States)

    Cai, Liangliang; Ye, Hongzhi; Yu, Fangrong; Li, Huiting; Chen, Jiashou; Liu, Xianxiang

    2013-05-01

    It has been recently shown that polysaccharides isolated from plants exhibit a number of beneficial therapeutic properties. Bauhinia championii (Benth.) Benth. has been widely used for the clinical treatment of knee osteoarthritis (OA) in China. However, the underlying molecular mechanisms of knee OA treatment have yet to be elucidated. In the present study, we investigated the effects of Bauhinia championii (Benth.) Benth. polysaccharides (BCBPs) on the proliferation and cell cycle of chondrocytes on 4-week-old male Sprague Dawley rats. Immunohistochemical staining was used to identify chondrocytes and an MTT assay was used to evaluate cell viability. Flow cytometry was used for cell cycle analysis. The mRNA and protein expression levels of cyclin D1, CDK4 and CDK6 in chondrocytes were detected using reverse transcription polymerase chain reaction (RT-PCR) and western blot analysis, respectively. The data demonstrate that BCBP treatment increased the viability of chondrocytes. In addition, BCBP treatment reduced the cell population in the G0/G1 phase, whereas the cell population was increased in the S phase. Furthermore, BCBP treatment enhanced the expression of cyclin D1, CDK4 and CDK6. These results indicate that BCBP treatment promotes cell proliferation by accelerating the G1/S transition.

  5. Topographic variation in redifferentiation capacity of chondrocytes in the adult human knee joint.

    Science.gov (United States)

    Stenhamre, H; Slynarski, K; Petrén, C; Tallheden, T; Lindahl, A

    2008-11-01

    The aim of this study was to investigate the topographic variation in matrix production and cell density in the adult human knee joint. Additionally, we have examined the redifferentiation potential of chondrocytes expanded in vitro from the different locations. Full thickness cartilage-bone biopsies were harvested from seven separate anatomical locations of healthy knee joints from deceased adult human donors. Chondrocytes were isolated, expanded in vitro and redifferentiated in a pellet mass culture. Biochemical analysis of total collagen, proteoglycans and cellular content as well as histology and immunohistochemistry were performed on biopsies and pellets. In the biochemical analysis of the biopsies, we found lower proteoglycan to collagen (GAG/HP) ratio in the non-weight bearing (NWB) areas compared to the weight bearing (WB) areas. The chondrocytes harvested from different locations in femur showed a significantly better attachment and proliferation ability as well as good post-expansion chondrogenic capacity in pellet mass culture compared with the cells harvested from tibia. These results demonstrate that there are differences in extra cellular content within the adult human knee in respect to GAG/HP ratio. Additionally, the data show that clear differences between chondrocytes harvested from femur and tibia from healthy human knee joints exist and that the differences are not completely abolished during the process of de- and redifferentiation. These findings emphasize the importance of the understanding of topographic variation in articular cartilage biology when approaching new cartilage repair strategies.

  6. Oxidative Stress Promotes Peroxiredoxin Hyperoxidation and Attenuates Pro-survival Signaling in Aging Chondrocytes*

    Science.gov (United States)

    Collins, John A.; Wood, Scott T.; Nelson, Kimberly J.; Rowe, Meredith A.; Carlson, Cathy S.; Chubinskaya, Susan; Poole, Leslie B.; Furdui, Cristina M.; Loeser, Richard F.

    2016-01-01

    Oxidative stress-mediated post-translational modifications of redox-sensitive proteins are postulated as a key mechanism underlying age-related cellular dysfunction and disease progression. Peroxiredoxins (PRX) are critical intracellular antioxidants that also regulate redox signaling events. Age-related osteoarthritis is a common form of arthritis that has been associated with mitochondrial dysfunction and oxidative stress. The objective of this study was to determine the effect of aging and oxidative stress on chondrocyte intracellular signaling, with a specific focus on oxidation of cytosolic PRX2 and mitochondrial PRX3. Menadione was used as a model to induce cellular oxidative stress. Compared with chondrocytes isolated from young adult humans, chondrocytes from older adults exhibited higher levels of PRX1–3 hyperoxidation basally and under conditions of oxidative stress. Peroxiredoxin hyperoxidation was associated with inhibition of pro-survival Akt signaling and stimulation of pro-death p38 signaling. These changes were prevented in cultured human chondrocytes by adenoviral expression of catalase targeted to the mitochondria (MCAT) and in cartilage explants from MCAT transgenic mice. Peroxiredoxin hyperoxidation was observed in situ in human cartilage sections from older adults and in osteoarthritic cartilage. MCAT transgenic mice exhibited less age-related osteoarthritis. These findings demonstrate that age-related oxidative stress can disrupt normal physiological signaling and contribute to osteoarthritis and suggest peroxiredoxin hyperoxidation as a potential mechanism. PMID:26797130

  7. Melatonin protects chondrocytes from impairment induced by glucocorticoids via NAD+-dependent SIRT1.

    Science.gov (United States)

    Yang, Wei; Kang, Xiaomin; Qin, Na; Li, Feng; Jin, Xinxin; Ma, Zhengmin; Qian, Zhuang; Wu, Shufang

    2017-10-01

    Intra-articular injection of glucocorticoids is used to relieve pain and inflammation in osteoarthritis patients, which is occasionally accompanied with the serious side effects of glucocorticoids in collagen-producing tissue. Melatonin is the major hormone released from the pineal gland and its beneficial effects on cartilage has been suggested. In the present study, we investigated the protective role of melatonin on matrix degeneration in chondrocytes induced by dexamethasone (Dex). The chondrocytes isolated from mice knee joint were treated with Dex, melatonin, EX527 and siRNA targeted for SIRT6, respectively. Dex treatment induced the loss of the extracellular matrix, NAD + /NADH ratio and NADPH concentration in chondrocytes. Melatonin alone have no effect on the quantity of proteoglycans and collagen type IIa1, however, the pretreatment of melatonin reversed the negative effects induced by Dex. Meanwhile, the significant decrease in NAD + /NADH ratio and NADPH concentration in Dex group were up-regulated by pretreatment of melatonin. Furthermore, it was revealed that inhibition of SIRT1 blocked the protective effects of melatonin. The enhancement of NAD + -dependent SIRT1 activity contributes to the chondroprotecfive effects of melatonin, which has a great benefit to prevent dexamethasone-induced chondrocytes impairment. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Effect of Collagen Type I or Type II on Chondrogenesis by Cultured Human Articular Chondrocytes

    NARCIS (Netherlands)

    Rutgers, M.; Saris, Daniël B.F.; Vonk, L.A.; van Rijen, M.H.P.; Akrum, V.; Langeveld, D.; van Boxtel, A.; Dhert, W.J.A.; Creemers, L.B.

    2013-01-01

    Introduction: Current cartilage repair procedures using autologous chondrocytes rely on a variety of carriers for implantation. Collagen types I and II are frequently used and valuable properties of both were shown earlier in vitro, although a preference for either was not demonstrated. Recently,

  9. Transforming growth factor-beta predominantly stimulates phenotypically changed chondrocytes in osteoarthritic human cartilage

    NARCIS (Netherlands)

    Lafeber, F. P.; van Roy, H. L.; van der Kraan, P. M.; van den Berg, W. B.; Bijlsma, J. W.

    1997-01-01

    One of the most prominent alterations that characterizes osteoarthritic cartilage damage is a reduction of proteoglycan content, reflecting an imbalance between synthesis and release of proteoglycans. Both synthesis and release depend on the activity of cartilage cells. Chondrocytes in the upper

  10. HIF-1α-induced HSP70 regulates anabolic responses in articular chondrocytes under hypoxic conditions.

    Science.gov (United States)

    Tsuchida, Shinji; Arai, Yuji; Takahashi, Kenji A; Kishida, Tsunao; Terauchi, Ryu; Honjo, Kuniaki; Nakagawa, Shuji; Inoue, Hiroaki; Ikoma, Kazuya; Ueshima, Keiichiro; Matsuki, Tomohiro; Mazda, Osam; Kubo, Toshikazu

    2014-08-01

    We assessed whether heat shock protein 70 (HSP70) is involved in hypoxia inducible factor 1 alpha (HIF-1α)-dependent anabolic pathways in articular chondrocytes under hypoxic conditions. Primary rabbit chondrocytes were cultured under normoxia (20% oxygen condition) or hypoxia (1% oxygen condition). Alternatively, cells cultured under normoxia were treated with CoCl2 , which induces HIF-1α, to simulate hypoxia, or transfected with siRNAs targeting HIF-1α (si-HIF-1α) and HSP70 (si-HSP70) under hypoxia. HSP70 expression was enhanced by the increased expression of HIF-1α under hypoxia or simulated hypoxia, but not in the presence of si-HIF-1α. Hypoxia-induced overexpression of ECM genes was significantly suppressed by si-HIF-1α or si-HSP70. Cell viability positively correlated with hypoxia, but transfection with si-HIF-1α or si-HSP70 abrogated the chondroprotective effects of hypoxia. Although LDH release from sodium nitroprusside-treated cells and the proportion of TUNEL positive cells were decreased under hypoxia, transfection with si-HIF-1α or si-HSP70 almost completely blocked these effects. These findings indicated that HIF-1α-induced HSP70 overexpression increased the expression levels of ECM genes and cell viability, and protected chondrocytes from apoptosis. HIF-1α may regulate the anabolic effects of chondrocytes under hypoxic conditions by regulating HSP70 expression. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  11. Effect of ceramic calcium-phosphorus ratio on chondrocyte-mediated biosynthesis and mineralization.

    Science.gov (United States)

    Boushell, Margaret K; Khanarian, Nora T; LeGeros, Raquel Z; Lu, Helen H

    2017-10-01

    The osteochondral interface functions as a structural barrier between cartilage and bone, maintaining tissue integrity postinjury and during homeostasis. Regeneration of this calcified cartilage region is thus essential for integrative cartilage healing, and hydrogel-ceramic composite scaffolds have been explored for calcified cartilage formation. The objective of this study is to test the hypothesis that Ca/P ratio of the ceramic phase of the composite scaffold regulates chondrocyte biosynthesis and mineralization potential. Specifically, the response of deep zone chondrocytes to two bioactive ceramics with different calcium-phosphorus ratios (1.35 ± 0.01 and 1.41 ± 0.02) was evaluated in agarose hydrogel scaffolds over two weeks in vitro. It was observed that the ceramic with higher calcium-phosphorus ratio enhanced chondrocyte proliferation, glycosaminoglycan production, and induced an early onset of alkaline phosphorus activity, while the ceramic with lower calcium-phosphorus ratio performed similarly to the ceramic-free control. These results underscore the importance of ceramic bioactivity in directing chondrocyte response, and demonstrate that Ca/P ratio is a key parameter to be considered in osteochondral scaffold design. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2694-2702, 2017. © 2017 Wiley Periodicals, Inc.

  12. Sprifermin (rhFGF18) enables proliferation of chondrocytes producing a hyaline cartilage matrix.

    Science.gov (United States)

    Gigout, A; Guehring, H; Froemel, D; Meurer, A; Ladel, C; Reker, D; Bay-Jensen, A C; Karsdal, M A; Lindemann, S

    2017-11-01

    Fibroblast growth factor (FGF) 18 has been shown to increase cartilage volume when injected intra-articularly in animal models of osteoarthritis (OA) and in patients with knee OA (during clinical development of the recombinant human FGF18, sprifermin). However, the exact nature of this effect is still unknown. In this study, we aimed to investigate the effects of sprifermin at the cellular level. A combination of different chondrocyte culture systems was used and the effects of sprifermin on proliferation, the phenotype and matrix production were evaluated. The involvement of MAPKs in sprifermin signalling was also studied. In monolayer, we observed that sprifermin promoted a round cell morphology and stimulated both cellular proliferation and Sox9 expression while strongly decreasing type I collagen expression. In 3D culture, sprifermin increased the number of matrix-producing chondrocytes, improved the type II:I collagen ratio and enabled human OA chondrocytes to produce a hyaline extracellular matrix (ECM). Furthermore, we found that sprifermin displayed a 'hit and run' mode of action, with intermittent exposure required for the compound to fully exert its anabolic effect. Finally, sprifermin appeared to signal through activation of ERK. Our results indicate that intermittent exposure to sprifermin leads to expansion of hyaline cartilage-producing chondrocytes. These in vitro findings are consistent with the increased cartilage volume observed in the knees of OA patients after intra-articular injection with sprifermin in clinical studies. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  13. Biochemical alterations in inflammatory reactive chondrocytes: evidence for intercellular network communication

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    Eva Skiöldebrand

    2018-01-01

    Full Text Available Chondrocytes are effectively involved in the pathophysiological processes of inflammation in joints. They form cellular processes in the superficial layer of the articular cartilage and form gap junction coupled syncytium to facilitate cell-to-cell communication. However, very little is known about their physiological cellular identity and communication. The aim with the present work is to evaluate the physiological behavior after stimulation with the inflammatory inducers interleukin-1β and lipopolysaccharide. The cytoskeleton integrity and intracellular Ca2+ release were assessed as indicators of inflammatory state. Cytoskeleton integrity was analyzed through cartilage oligomeric matrix protein and actin labeling with an Alexa 488-conjugated phalloidin probe. Ca2+ responses were assessed through the Ca2+ sensitive fluorophore Fura-2/AM. Western blot analyses of several inflammatory markers were performed. The results show reorganization of the actin filaments. Glutamate, 5-hydoxytryptamine, and ATP evoked intracellular Ca2+ release changed from single peaks to oscillations after inflammatory induction in the chondrocytes. The expression of toll-like receptor 4, the glutamate transporters GLAST and GLT-1, and the matrix metalloproteinase-13 increased. This work demonstrates that chondrocytes are a key part in conditions that lead to inflammation in the cartilage. The inflammatory inducers modulate the cytoskeleton, the Ca2+ signaling, and several inflammatory parameters. In conclusion, our data show that the cellular responses to inflammatory insults from healthy and inflammatory chondrocytes resemble those previously observed in astrocyte and cardiac fibroblasts networks.

  14. Latexin is involved in bone morphogenetic protein-2-induced chondrocyte differentiation

    International Nuclear Information System (INIS)

    Kadouchi, Ichiro; Sakamoto, Kei; Tangjiao, Liu; Murakami, Takashi; Kobayashi, Eiji; Hoshino, Yuichi; Yamaguchi, Akira

    2009-01-01

    Latexin is the only known carboxypeptidase A inhibitor in mammals. We previously demonstrated that BMP-2 significantly induced latexin expression in Runx2-deficient mesenchymal cells (RD-C6 cells), during chondrocyte and osteoblast differentiation. In this study, we investigated latexin expression in the skeleton and its role in chondrocyte differentiation. Immunohistochemical studies revealed that proliferating and prehypertrophic chondrocytes expressed latexin during skeletogenesis and bone fracture repair. In the early phase of bone fracture, latexin mRNA expression was dramatically upregulated. BMP-2 upregulated the expression of the mRNAs of latexin, Col2a1, and the gene encoding aggrecan (Agc1) in a micromass culture of C3H10T1/2 cells. Overexpression of latexin additively stimulated the BMP-2-induced expression of the mRNAs of Col2a, Agc1, and Col10a1. BMP-2 treatment upregulated Sox9 expression, and Sox9 stimulated the promoter activity of latexin. These results indicate that latexin is involved in BMP-2-induced chondrocyte differentiation and plays an important role in skeletogenesis and skeletal regeneration.

  15. Characteristic complications after autologous chondrocyte implantation for cartilage defects of the knee joint

    DEFF Research Database (Denmark)

    Niemeyer, Philipp; Pestka, Jan M; Kreuz, Peter C

    2008-01-01

    BACKGROUND: Although autologous chondrocyte implantation (ACI) is a well-established therapy for the treatment of isolated cartilage defects of the knee joint, little is known about typical complications and their treatment after ACI. HYPOTHESIS: Unsatisfactory outcome after ACI is associated...

  16. Acquiring Chondrocyte Phenotype from Human Mesenchymal Stem Cells under Inflammatory Conditions

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    Masahiro Kondo

    2014-11-01

    Full Text Available An inflammatory milieu breaks down the cartilage matrix and induces chondrocyte apoptosis, resulting in cartilage destruction in patients with cartilage degenerative diseases, such as rheumatoid arthritis or osteoarthritis. Because of the limited regenerative ability of chondrocytes, defects in cartilage are irreversible and difficult to repair. Mesenchymal stem cells (MSCs are expected to be a new tool for cartilage repair because they are present in the cartilage and are able to differentiate into multiple lineages of cells, including chondrocytes. Although clinical trials using MSCs for patients with cartilage defects have already begun, its efficacy and repair mechanisms remain unknown. A PubMed search conducted in October 2014 using the following medical subject headings (MeSH terms: mesenchymal stromal cells, chondrogenesis, and cytokines resulted in 204 articles. The titles and abstracts were screened and nine articles relevant to “inflammatory” cytokines and “human” MSCs were identified. Herein, we review the cell biology and mechanisms of chondrocyte phenotype acquisition from human MSCs in an inflammatory milieu and discuss the clinical potential of MSCs for cartilage repair.

  17. Upregulation of matrix synthesis in chondrocyte-seeded agarose following sustained bi-axial cyclic loading

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    Belinda Pingguan-Murphy

    2012-08-01

    Full Text Available OBJECTIVES: The promotion of extracellular matrix synthesis by chondrocytes is a requisite part of an effective cartilage tissue engineering strategy. The aim of this in vitro study was to determine the effect of bi-axial cyclic mechanical loading on cell proliferation and the synthesis of glycosaminoglycans by chondrocytes in threedimensional cultures. METHOD: A strain comprising 10% direct compression and 1% compressive shear was applied to bovine chondrocytes seeded in an agarose gel during two 12-hour conditioning periods separated by a 12-hour resting period. RESULTS: The bi-axial-loaded chondrocytes demonstrated a significant increase in glycosaminoglycan synthesis compared with samples exposed to uni-axial or no loading over the same period (p<0.05. The use of a free-swelling recovery period prior to the loading regime resulted in additional glycosaminoglycan production and a significant increase in DNA content (p<0.05, indicating cell proliferation. CONCLUSIONS: These results demonstrate that the use of a bi-axial loading regime results in increased matrix production compared with uni-axial loading.

  18. Xiphoid Process-Derived Chondrocytes: A Novel Cell Source for Elastic Cartilage Regeneration

    Science.gov (United States)

    Nam, Seungwoo; Cho, Wheemoon; Cho, Hyunji; Lee, Jungsun

    2014-01-01

    Reconstruction of elastic cartilage requires a source of chondrocytes that display a reliable differentiation tendency. Predetermined tissue progenitor cells are ideal candidates for meeting this need; however, it is difficult to obtain donor elastic cartilage tissue because most elastic cartilage serves important functions or forms external structures, making these tissues indispensable. We found vestigial cartilage tissue in xiphoid processes and characterized it as hyaline cartilage in the proximal region and elastic cartilage in the distal region. Xiphoid process-derived chondrocytes (XCs) showed superb in vitro expansion ability based on colony-forming unit fibroblast assays, cell yield, and cumulative cell growth. On induction of differentiation into mesenchymal lineages, XCs showed a strong tendency toward chondrogenic differentiation. An examination of the tissue-specific regeneration capacity of XCs in a subcutaneous-transplantation model and autologous chondrocyte implantation model confirmed reliable regeneration of elastic cartilage regardless of the implantation environment. On the basis of these observations, we conclude that xiphoid process cartilage, the only elastic cartilage tissue source that can be obtained without destroying external shape or function, is a source of elastic chondrocytes that show superb in vitro expansion and reliable differentiation capacity. These findings indicate that XCs could be a valuable cell source for reconstruction of elastic cartilage. PMID:25205841

  19. Chondrogenic differentiation of human articular chondrocytes differs in biodegradable PGA/PLA scaffolds

    DEFF Research Database (Denmark)

    Zwingmann, Joern; Mehlhorn, Alexander T; Südkamp, Norbert

    2007-01-01

    Cartilage tissue engineering is applied clinically to cover and regenerate articular cartilage defects. Two bioresorbable nonwoven scaffolds, polyglycolic acid (PGA) and poly(lactic-co-glycolic acid) (PLGA) (90/10 copolymer of L-lactide and glycolide), were seeded with human chondrocytes after in...

  20. CCN2/CTGF is required for matrix organization and to protect growth plate chondrocytes from cellular stress.

    Science.gov (United States)

    Hall-Glenn, Faith; Aivazi, Armen; Akopyan, Lusi; Ong, Jessica R; Baxter, Ruth R; Benya, Paul D; Goldschmeding, Roel; van Nieuwenhoven, Frans A; Hunziker, Ernst B; Lyons, Karen M

    2013-08-01

    CCN2 (connective tissue growth factor (CTGF/CCN2)) is a matricellular protein that utilizes integrins to regulate cell proliferation, migration and survival. The loss of CCN2 leads to perinatal lethality resulting from a severe chondrodysplasia. Upon closer inspection of Ccn2 mutant mice, we observed defects in extracellular matrix (ECM) organization and hypothesized that the severe chondrodysplasia caused by loss of CCN2 might be associated with defective chondrocyte survival. Ccn2 mutant growth plate chondrocytes exhibited enlarged endoplasmic reticula (ER), suggesting cellular stress. Immunofluorescence analysis confirmed elevated stress in Ccn2 mutants, with reduced stress observed in Ccn2 overexpressing transgenic mice. In vitro studies revealed that Ccn2 is a stress responsive gene in chondrocytes. The elevated stress observed in Ccn2-/- chondrocytes is direct and mediated in part through integrin α5. The expression of the survival marker NFκB and components of the autophagy pathway were decreased in Ccn2 mutant growth plates, suggesting that CCN2 may be involved in mediating chondrocyte survival. These data demonstrate that absence of a matricellular protein can result in increased cellular stress and highlight a novel protective role for CCN2 in chondrocyte survival. The severe chondrodysplasia caused by the loss of CCN2 may be due to increased chondrocyte stress and defective activation of autophagy pathways, leading to decreased cellular survival. These effects may be mediated through nuclear factor κB (NFκB) as part of a CCN2/integrin/NFκB signaling cascade.

  1. A Tumor Suppressor Gene Product, Platelet-Derived Growth Factor Receptor-Like Protein Controls Chondrocyte Proliferation and Differentiation.

    Science.gov (United States)

    Kawata, Kazumi; Kubota, Satoshi; Eguchi, Takanori; Aoyama, Eriko; Moritani, Norifumi H; Oka, Morihiko; Kawaki, Harumi; Takigawa, Masaharu

    2017-11-01

    The platelet-derived growth factor receptor-like (PDGFRL) gene is regarded as a tumor suppressor gene. However, nothing is known about the molecular function of PDGFRL. In this study, we initially clarified its function in chondrocytes. Among all cell lines examined, the PDGFRL mRNA level was the highest in chondrocytic HCS-2/8 cells. Interestingly, the proliferation of chondrocytic HCS-2/8 cells was promoted by PDGFRL overexpression, whereas that of the breast cancer-derived MDA-MB-231 cells was inhibited. Of note, in PDGFRL-overexpressing HCS-2/8 cells, the expression of chondrocyte differentiation marker genes, SOX9, ACAN, COL2A1, COL10A1, and ALP, was decreased. Moreover, we confirmed the expression of PDGFRL mRNA in normal cartilage tissue and chondrocytes. Eventually, the expression of PDGFRL mRNA in condrocytes except in the case of hypertrophic chondrocytes was demonstrated in vivo and in vitro. These findings suggest that PDGFRL plays the different roles, depending upon cell types. Particularly, in chondrocytes, PDGFRL may play a new and important role which is distinct from the function previously reported. J. Cell. Biochem. 118: 4033-4044, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  2. Lithium Chloride Dependent Glycogen Synthase Kinase 3 Inactivation Links Oxidative DNA Damage, Hypertrophy and Senescence in Human Articular Chondrocytes and Reproduces Chondrocyte Phenotype of Obese Osteoarthritis Patients.

    Directory of Open Access Journals (Sweden)

    Serena Guidotti

    Full Text Available Recent evidence suggests that GSK3 activity is chondroprotective in osteoarthritis (OA, but at the same time, its inactivation has been proposed as an anti-inflammatory therapeutic option. Here we evaluated the extent of GSK3β inactivation in vivo in OA knee cartilage and the molecular events downstream GSK3β inactivation in vitro to assess their contribution to cell senescence and hypertrophy.In vivo level of phosphorylated GSK3β was analyzed in cartilage and oxidative damage was assessed by 8-oxo-deoxyguanosine staining. The in vitro effects of GSK3β inactivation (using either LiCl or SB216763 were evaluated on proliferating primary human chondrocytes by combined confocal microscopy analysis of Mitotracker staining and reactive oxygen species (ROS production (2',7'-dichlorofluorescin diacetate staining. Downstream effects on DNA damage and senescence were investigated by western blot (γH2AX, GADD45β and p21, flow cytometric analysis of cell cycle and light scattering properties, quantitative assessment of senescence associated β galactosidase activity, and PAS staining.In vivo chondrocytes from obese OA patients showed higher levels of phosphorylated GSK3β, oxidative damage and expression of GADD45β and p21, in comparison with chondrocytes of nonobese OA patients. LiCl mediated GSK3β inactivation in vitro resulted in increased mitochondrial ROS production, responsible for reduced cell proliferation, S phase transient arrest, and increase in cell senescence, size and granularity. Collectively, western blot data supported the occurrence of a DNA damage response leading to cellular senescence with increase in γH2AX, GADD45β and p21. Moreover, LiCl boosted 8-oxo-dG staining, expression of IKKα and MMP-10.In articular chondrocytes, GSK3β activity is required for the maintenance of proliferative potential and phenotype. Conversely, GSK3β inactivation, although preserving chondrocyte survival, results in functional impairment via

  3. Lithium Chloride Dependent Glycogen Synthase Kinase 3 Inactivation Links Oxidative DNA Damage, Hypertrophy and Senescence in Human Articular Chondrocytes and Reproduces Chondrocyte Phenotype of Obese Osteoarthritis Patients.

    Science.gov (United States)

    Guidotti, Serena; Minguzzi, Manuela; Platano, Daniela; Cattini, Luca; Trisolino, Giovanni; Mariani, Erminia; Borzì, Rosa Maria

    2015-01-01

    Recent evidence suggests that GSK3 activity is chondroprotective in osteoarthritis (OA), but at the same time, its inactivation has been proposed as an anti-inflammatory therapeutic option. Here we evaluated the extent of GSK3β inactivation in vivo in OA knee cartilage and the molecular events downstream GSK3β inactivation in vitro to assess their contribution to cell senescence and hypertrophy. In vivo level of phosphorylated GSK3β was analyzed in cartilage and oxidative damage was assessed by 8-oxo-deoxyguanosine staining. The in vitro effects of GSK3β inactivation (using either LiCl or SB216763) were evaluated on proliferating primary human chondrocytes by combined confocal microscopy analysis of Mitotracker staining and reactive oxygen species (ROS) production (2',7'-dichlorofluorescin diacetate staining). Downstream effects on DNA damage and senescence were investigated by western blot (γH2AX, GADD45β and p21), flow cytometric analysis of cell cycle and light scattering properties, quantitative assessment of senescence associated β galactosidase activity, and PAS staining. In vivo chondrocytes from obese OA patients showed higher levels of phosphorylated GSK3β, oxidative damage and expression of GADD45β and p21, in comparison with chondrocytes of nonobese OA patients. LiCl mediated GSK3β inactivation in vitro resulted in increased mitochondrial ROS production, responsible for reduced cell proliferation, S phase transient arrest, and increase in cell senescence, size and granularity. Collectively, western blot data supported the occurrence of a DNA damage response leading to cellular senescence with increase in γH2AX, GADD45β and p21. Moreover, LiCl boosted 8-oxo-dG staining, expression of IKKα and MMP-10. In articular chondrocytes, GSK3β activity is required for the maintenance of proliferative potential and phenotype. Conversely, GSK3β inactivation, although preserving chondrocyte survival, results in functional impairment via induction of

  4. Bauhinia championi (Benth.) Benth. polysaccharides upregulate Wnt/β-catenin signaling in chondrocytes.

    Science.gov (United States)

    Li, Huiting; Li, Xihai; Liu, Guozhong; Chen, Jiashou; Weng, Xiaping; Liu, Fayuan; Xu, Huifeng; Liu, Xianxiang; Ye, Hongzhi

    2013-12-01

    Bauhinia championi (Benth.) Benth. polysaccharides (BCBPs), extracted from Bauhinia championi (Benth.) Benth., which has been used in traditional Chinese medicine (TCM) for the treatment of osteoarthritis (OA), are the bioactive constituents of Bauhinia championi (Benth.) rattan. However, the molecular mechanisms responsible for their effects on OA are poorly understood. The Wnt/β-catenin signaling pathway plays an important role in the proliferation of chondrocytes. In the present study, the effects of BCBPs on Wnt/β-catenin signaling in chondrocytes were investigated. BCBPs were obtained by hot-water extraction and identified by the modified high performance liquid chromatography (HPLC) method. Chondrocytes were isolated from the knees of Sprague‑Dawley rats and identified by type II collagen immunohistochemistry. The chondrocytes were treated with or without BCBPs for 48 h. Cell viability was evaluated by MTT assay. The mRNA and protein levels of Wnt-4, β-catenin, Frizzled-2, glycogen synthase kinase (GSK)-3β, cyclin D1 and collagen II were detected by western blot analysis and reverse transcription PCR (RT-PCR), respectively. We found that the BCBPs contained at least seven monosaccharides, including D-mannose, rhamnose, D-(+) glucuronic acid, D-(+) galacturonic acid, D-glucose, galactose and arabinose. The cell viability of the chondrocytes treated with 50, 100 and 200 µg/ml BCBPs was significantly higher than that of the chondroctyes in the control group (treated with 0 µg/ml BCBPs). Furthermore, compared with the control group, the mRNA and protein expression of Wnt-4, β-catenin, Frizzled-2 and cyclin D1 in the BCBP-treated groups markedly increased, whereas the mRNA and protein expression of GSK-3β significantly decreased. Of note, the dose of 100 µg/ml BCBPs was more effective than the dose of 50 µg/ml BCBPs and 200 µg/ml BCBPs. In addition, we found that treatment with BCBPs upregulated the protein levels of collagen II in the

  5. Culture temperature affects human chondrocyte messenger RNA expression in monolayer and pellet culture systems.

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    Akira Ito

    Full Text Available Cell-based therapy has been explored for articular cartilage regeneration. Autologous chondrocyte implantation is a promising cell-based technique for repairing articular cartilage defects. However, there are several issues such as chondrocyte de-differentiation. While numerous studies have been designed to overcome some of these issues, only a few have focused on the thermal environment that can affect chondrocyte metabolism and phenotype. In this study, the effects of different culture temperatures on human chondrocyte metabolism- and phenotype-related gene expression were investigated in 2D and 3D environments. Human chondrocytes were cultured in a monolayer or in a pellet culture system at three different culture temperatures (32°C, 37°C, and 41°C for 3 days. The results showed that the total RNA level, normalized to the threshold cycle value of internal reference genes, was higher at lower temperatures in both culture systems. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH and citrate synthase (CS, which are involved in glycolysis and the citric acid cycle, respectively, were expressed at similar levels at 32°C and 37°C in pellet cultures, but the levels were significantly lower at 41°C. Expression of the chondrogenic markers, collagen type IIA1 (COL2A1 and aggrecan (ACAN, was higher at 37°C than at 32°C and 41°C in both culture systems. However, this phenomenon did not coincide with SRY (sex-determining region Y-box 9 (SOX9, which is a fundamental transcription factor for chondrogenesis, indicating that a SOX9-independent pathway might be involved in this phenomenon. In conclusion, the expression of chondrocyte metabolism-related genes at 32°C was maintained or enhanced compared to that at 37°C. However, chondrogenesis-related genes were further induced at 37°C in both culture systems. Therefore, manipulating the culture temperature may be an advantageous approach for regulating human chondrocyte metabolic activity and

  6. Culture temperature affects human chondrocyte messenger RNA expression in monolayer and pellet culture systems.

    Science.gov (United States)

    Ito, Akira; Nagai, Momoko; Tajino, Junichi; Yamaguchi, Shoki; Iijima, Hirotaka; Zhang, Xiangkai; Aoyama, Tomoki; Kuroki, Hiroshi

    2015-01-01

    Cell-based therapy has been explored for articular cartilage regeneration. Autologous chondrocyte implantation is a promising cell-based technique for repairing articular cartilage defects. However, there are several issues such as chondrocyte de-differentiation. While numerous studies have been designed to overcome some of these issues, only a few have focused on the thermal environment that can affect chondrocyte metabolism and phenotype. In this study, the effects of different culture temperatures on human chondrocyte metabolism- and phenotype-related gene expression were investigated in 2D and 3D environments. Human chondrocytes were cultured in a monolayer or in a pellet culture system at three different culture temperatures (32°C, 37°C, and 41°C) for 3 days. The results showed that the total RNA level, normalized to the threshold cycle value of internal reference genes, was higher at lower temperatures in both culture systems. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and citrate synthase (CS), which are involved in glycolysis and the citric acid cycle, respectively, were expressed at similar levels at 32°C and 37°C in pellet cultures, but the levels were significantly lower at 41°C. Expression of the chondrogenic markers, collagen type IIA1 (COL2A1) and aggrecan (ACAN), was higher at 37°C than at 32°C and 41°C in both culture systems. However, this phenomenon did not coincide with SRY (sex-determining region Y)-box 9 (SOX9), which is a fundamental transcription factor for chondrogenesis, indicating that a SOX9-independent pathway might be involved in this phenomenon. In conclusion, the expression of chondrocyte metabolism-related genes at 32°C was maintained or enhanced compared to that at 37°C. However, chondrogenesis-related genes were further induced at 37°C in both culture systems. Therefore, manipulating the culture temperature may be an advantageous approach for regulating human chondrocyte metabolic activity and chondrogenesis.

  7. Reconstruction of Hyaline Cartilage Deep Layer Properties in 3-Dimensional Cultures of Human Articular Chondrocytes.

    Science.gov (United States)

    Nanduri, Vibudha; Tattikota, Surendra Mohan; T, Avinash Raj; Sriramagiri, Vijaya Rama Rao; Kantipudi, Suma; Pande, Gopal

    2014-06-01

    Articular cartilage (AC) injuries and malformations are commonly noticed because of trauma or age-related degeneration. Many methods have been adopted for replacing or repairing the damaged tissue. Currently available AC repair methods, in several cases, fail to yield good-quality long-lasting results, perhaps because the reconstructed tissue lacks the cellular and matrix properties seen in hyaline cartilage (HC). To reconstruct HC tissue from 2-dimensional (2D) and 3-dimensional (3D) cultures of AC-derived human chondrocytes that would specifically exhibit the cellular and biochemical properties of the deep layer of HC. Descriptive laboratory study. Two-dimensional cultures of human AC-derived chondrocytes were established in classical medium (CM) and newly defined medium (NDM) and maintained for a period of 6 weeks. These cells were suspended in 2 mm-thick collagen I gels, placed in 24-well culture inserts, and further cultured up to 30 days. Properties of chondrocytes, grown in 2D cultures and the reconstructed 3D cartilage tissue, were studied by optical and scanning electron microscopic techniques, immunohistochemistry, and cartilage-specific gene expression profiling by reverse transcription polymerase chain reaction and were compared with those of the deep layer of native human AC. Two-dimensional chondrocyte cultures grown in NDM, in comparison with those grown in CM, showed more chondrocyte-specific gene activity and matrix properties. The NDM-grown chondrocytes in 3D cultures also showed better reproduction of deep layer properties of HC, as confirmed by microscopic and gene expression analysis. The method used in this study can yield cartilage tissue up to approximately 1.6 cm in diameter and 2 mm in thickness that satisfies the very low cell density and matrix composition properties present in the deep layer of normal HC. This study presents a novel and reproducible method for long-term culture of AC-derived chondrocytes and reconstruction of cartilage

  8. Increased adipogenesis in cultured embryonic chondrocytes and in adult bone marrow of dominant negative Erg transgenic mice.

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    Sébastien Flajollet

    Full Text Available In monolayer culture, primary articular chondrocytes have an intrinsic tendency to lose their phenotype during expansion. The molecular events underlying this chondrocyte dedifferentiation are still largely unknown. Several transcription factors are important for chondrocyte differentiation. The Ets transcription factor family may be involved in skeletal development. One family member, the Erg gene, is mainly expressed during cartilage formation. To further investigate the potential role of Erg in the maintenance of the chondrocyte phenotype, we isolated and cultured chondrocytes from the rib cartilage of embryos of transgenic mice that express a dominant negative form of Erg (DN-Erg during cartilage formation. DN-Erg expression in chondrocytes cultured for up to 20 days did not affect the early dedifferentiation usually observed in cultured chondrocytes. However, lipid droplets accumulated in DN-Erg chondrocytes, suggesting adipocyte emergence. Transcriptomic analysis using a DNA microarray, validated by quantitative RT-PCR, revealed strong differential gene expression, with a decrease in chondrogenesis-related markers and an increase in adipogenesis-related gene expression in cultured DN-Erg chondrocytes. These results indicate that Erg is involved in either maintaining the chondrogenic phenotype in vitro or in cell fate orientation. Along with the in vitro studies, we compared adipocyte presence in wild-type and transgenic mice skeletons. Histological investigations revealed an increase in the number of adipocytes in the bone marrow of adult DN-Erg mice even though no adipocytes were detected in embryonic cartilage or bone. These findings suggest that the Ets transcription factor family may contribute to the homeostatic balance in skeleton cell plasticity.

  9. Long Intergenic Noncoding RNAs Mediate the Human Chondrocyte Inflammatory Response and Are Differentially Expressed in Osteoarthritis Cartilage.

    Science.gov (United States)

    Pearson, Mark J; Philp, Ashleigh M; Heward, James A; Roux, Benoit T; Walsh, David A; Davis, Edward T; Lindsay, Mark A; Jones, Simon W

    2016-04-01

    To identify long noncoding RNAs (lncRNAs), including long intergenic noncoding RNAs (lincRNAs), antisense RNAs, and pseudogenes, associated with the inflammatory response in human primary osteoarthritis (OA) chondrocytes and to explore their expression and function in OA. OA cartilage was obtained from patients with hip or knee OA following joint replacement surgery. Non-OA cartilage was obtained from postmortem donors and patients with fracture of the neck of the femur. Primary OA chondrocytes were isolated by collagenase digestion. LncRNA expression analysis was performed by RNA sequencing (RNAseq) and quantitative reverse transcriptase-polymerase chain reaction. Modulation of lncRNA chondrocyte expression was achieved using LNA longRNA GapmeRs (Exiqon). Cytokine production was measured with Luminex. RNAseq identified 983 lncRNAs in primary human hip OA chondrocytes, 183 of which had not previously been identified. Following interleukin-1β (IL-1β) stimulation, we identified 125 lincRNAs that were differentially expressed. The lincRNA p50-associated cyclooxygenase 2-extragenic RNA (PACER) and 2 novel chondrocyte inflammation-associated lincRNAs (CILinc01 and CILinc02) were differentially expressed in both knee and hip OA cartilage compared to non-OA cartilage. In primary OA chondrocytes, these lincRNAs were rapidly and transiently induced in response to multiple proinflammatory cytokines. Knockdown of CILinc01 and CILinc02 expression in human chondrocytes significantly enhanced the IL-1-stimulated secretion of proinflammatory cytokines. The inflammatory response in human OA chondrocytes is associated with widespread changes in the profile of lncRNAs, including PACER, CILinc01, and CILinc02. Differential expression of CILinc01 and CIinc02 in hip and knee OA cartilage, and their role in modulating cytokine production during the chondrocyte inflammatory response, suggest that they may play an important role in mediating inflammation-driven cartilage degeneration in

  10. Indian hedgehog signaling promotes chondrocyte differentiation in enchondral ossification in human cervical ossification of the posterior longitudinal ligament.

    Science.gov (United States)

    Sugita, Daisuke; Yayama, Takafumi; Uchida, Kenzo; Kokubo, Yasuo; Nakajima, Hideaki; Yamagishi, Atsushi; Takeura, Naoto; Baba, Hisatoshi

    2013-10-15

    Histological, immunohistochemical, and immunoblot analyses of the expression of Indian hedgehog (Ihh) signaling in human cervical ossification of the posterior longitudinal ligament (OPLL). To examine the hypothesis that Ihh signaling in correlation with Sox9 and parathyroid-related peptide hormone (PTHrP) facilitates chondrocyte differentiation in enchondral ossification process in human cervical OPLL. In enchondral ossification, certain transcriptional factors regulate cell differentiation. OPLL is characterized by overexpression of these factors and disturbance of the normal cell differentiation process. Ihh signaling is essential for enchondral ossification, especially in chondrocyte hypertrophy. Samples of ossified ligaments were harvested from 45 patients who underwent anterior cervical decompressive surgery for symptomatic OPLL, and 6 control samples from patients with cervical spondylotic myelopathy/radiculopathy without OPLL. The harvested sections were stained with hematoxylin-eosin and toluidine blue, examined by transmission electron microscopy, and immunohistochemically stained for Ihh, PTHrP, Sox9, type X, XI collagen, and alkaline phosphatase. Immunoblot analysis was performed in cultured cells derived from the posterior longitudinal ligaments in the vicinity of the ossified plaque and examined for the expression of these factors. The ossification front in OPLL contained chondrocytes at various differentiation stages, including proliferating chondrocytes in fibrocartilaginous area, hypertrophic chondrocytes around the calcification front, and apoptotic chondrocytes near the ossified area. Immunoreactivity for Ihh and Sox9 was evident in proliferating chondrocytes and was strongly positive for PTHrP in hypertrophic chondrocytes. Mesenchymal cells with blood vessel formation were positive for Ihh, PTHrP, and Sox9. Cultured cells from OPLL tissues expressed significantly higher levels of Ihh, PTHrP, and Sox9 than those in non-OPLL cells. Our results

  11. Effect of chondrocyte-derived early extracellular matrix on chondrogenesis of placenta-derived mesenchymal stem cells.

    Science.gov (United States)

    Park, Yong-Beom; Seo, Sinji; Kim, Jin-A; Heo, Jin-Chul; Lim, Young-Cheol; Ha, Chul-Won

    2015-06-24

    The extracellular matrix (ECM) surrounding cells contains a variety of proteins that provide structural support and regulate cellular functions. Previous studies have shown that decellularized ECM isolated from tissues or cultured cells can be used to improve cell differentiation in tissue engineering applications. In this study we evaluated the effect of decellularized chondrocyte-derived ECM (CDECM) on the chondrogenesis of human placenta-derived mesenchymal stem cells (hPDMSCs) in a pellet culture system. After incubation with or without chondrocyte-derived ECM in chondrogenic medium for 1 or 3 weeks, the sizes and wet masses of the cell pellets were compared with untreated controls (hPDMSCs incubated in chondrogenic medium without chondrocyte-derived ECM). In addition, histologic analysis of the cell pellets (Safranin O and collagen type II staining) and quantitative reverse transcription-PCR analysis of chondrogenic markers (aggrecan, collagen type II, and SOX9) were carried out. Our results showed that the sizes and masses of hPDMSC pellets incubated with chondrocyte-derived ECM were significantly higher than those of untreated controls. Differentiation of hPDMSCs (both with and without chondrocyte-derived ECM) was confirmed by Safranin O and collagen type II staining. Chondrogenic marker expression and glycosaminoglycan (GAG) levels were significantly higher in hPDMSC pellets incubated with chondrocyte-derived ECM compared with untreated controls, especially in cells precultured with chondrocyte-derived ECM for 7 d. Taken together, these results demonstrate that chondrocyte-derived ECM enhances the chondrogenesis of hPDMSCs, and this effect is further increased by preculture with chondrocyte-derived ECM. This preculture method for hPDMSC chondrogenesis represents a promising approach for cartilage tissue engineering.

  12. The identification of CD163 expressing phagocytic chondrocytes in joint cartilage and its novel scavenger role in cartilage degradation.

    Directory of Open Access Journals (Sweden)

    Kai Jiao

    Full Text Available BACKGROUND: Cartilage degradation is a typical characteristic of arthritis. This study examined whether there was a subset of phagocytic chondrocytes that expressed the specific macrophage marker, CD163, and investigated their role in cartilage degradation. METHODS: Cartilage from the knee and temporomandibular joints of Sprague-Dawley rats was harvested. Cartilage degradation was experimentally-induced in rat temporomandibular joints, using published biomechanical dental methods. The expression levels of CD163 and inflammatory factors within cartilage, and the ability of CD163(+ chondrocytes to conduct phagocytosis were investigated. Cartilage from the knees of patients with osteoarthritis and normal cartilage from knee amputations was also investigated. RESULTS: In the experimentally-induced degrading cartilage from temporomandibular joints, phagocytes were capable of engulfing neighboring apoptotic and necrotic cells, and the levels of CD163, TNF-α and MMPs were all increased (P0.05. CD163(+ chondrocytes were found in the cartilage mid-zone of temporomandibular joints and knee from healthy, three-week old rats. Furthermore, an increased number of CD163(+ chondrocytes with enhanced phagocytic activity were present in Col-II(+ chondrocytes isolated from the degraded cartilage of temporomandibular joints in the eight-week experimental group compared with their age-matched controls. Increased number with enhanced phagocytic activity of CD163(+ chondrocytes were also found in isolated Col-II(+ chondrocytes stimulated with TNF-α (P<0.05. Mid-zone distribution of CD163(+ cells accompanied with increased expression of CD163 and TNF-α were further confirmed in the isolated Col-II(+ chondrocytes from the knee cartilage of human patients with osteoarthritis, in contrast to the controls (both P<0.05. CONCLUSIONS: An increased number of CD163(+ chondrocytes with enhanced phagocytic activity were discovered within degraded joint cartilage, indicating a

  13. Cartilage grafting in nasal reconstruction.

    Science.gov (United States)

    Immerman, Sara; White, W Matthew; Constantinides, Minas

    2011-02-01

    Nasal reconstruction after resection for cutaneous malignancies poses a unique challenge to facial plastic surgeons. The nose, a unique 3-D structure, not only must remain functional but also be aesthetically pleasing to patients. A complete understanding of all the layers of the nose and knowledge of available cartilage grafting material is necessary. Autogenous material, namely septal, auricular, and costal cartilage, is the most favored material in a free cartilage graft or a composite cartilage graft. All types of material have advantages and disadvantages that should guide the most appropriate selection to maximize the functional and cosmetic outcomes for patients. Copyright © 2011 Elsevier Inc. All rights reserved.

  14. Inheritance of graft compatibility in Douglas fir.

    Science.gov (United States)

    D.L. Copes

    1973-01-01

    Graft compatibility of genetically related and unrelated rootstock-scion combinations was compared. Scion clones were 75% compatible when grafted on half-related rootstocks but only 56% compatible when grafted on unrelated rootstocks. Most variance associated with graft incompatibility in Douglas-fir appears to be caused by multiple genes.

  15. Graft union formation in Douglas-fir.

    Science.gov (United States)

    D.L. Copes

    1969-01-01

    Greenhouse-grown Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco) graft unions were examined between 2 and 84 days after grafting. Room temperature was maintained at 60-70 F throughout the growing season. In most respects grafts of Douglas-fir followed development patterns previously reported for spruce and pine grafts, but specific differences...

  16. Articular chondrocyte network mediated by gap junctions: role in metabolic cartilage homeostasis

    Science.gov (United States)

    Mayan, Maria D; Gago-Fuentes, Raquel; Carpintero-Fernandez, Paula; Fernandez-Puente, Patricia; Filgueira-Fernandez, Purificacion; Goyanes, Noa; Valiunas, Virginijus; Brink, Peter R; Goldberg, Gary S; Blanco, Francisco J

    2017-01-01

    Objective This study investigated whether chondrocytes within the cartilage matrix have the capacity to communicate through intercellular connections mediated by voltage-gated gap junction (GJ) channels. Methods Frozen cartilage samples were used for immunofluorescence and immunohistochemistry assays. Samples were embedded in cacodylate buffer before dehydration for scanning electron microscopy. Co-immunoprecipitation experiments and mass spectrometry (MS) were performed to identify proteins that interact with the C-terminal end of Cx43. GJ communication was studied through in situ electroporation, electrophysiology and dye injection experiments. A transwell layered culture system and MS were used to identify and quantify transferred amino acids. Results Microscopic images revealed the presence of multiple cellular projections connecting chondrocytes within the matrix. These projections were between 5 and 150 μm in length. MS data analysis indicated that the C-terminus of Cx43 interacts with several cytoskeletal proteins implicated in Cx trafficking and GJ assembly, including α-tubulin and β-tubulin, actin, and vinculin. Electrophysiology experiments demonstrated that 12-mer oligonucleotides could be transferred between chondrocytes within 12 min after injection. Glucose was homogeneously distributed within 22 and 35 min. No transfer was detected when glucose was electroporated into A549 cells, which have no GJs. Transwell layered culture systems coupled with MS analysis revealed connexins can mediate the transfer of L-lysine and L-arginine between chondrocytes. Conclusions This study reveals that intercellular connections between chondrocytes contain GJs that play a key role in cell-cell communication and a metabolic function by exchange of nutrients including glucose and essential amino acids. A three-dimensional cellular network mediated through GJs might mediate metabolic and physiological homeostasis to maintain cartilage tissue. PMID:24225059

  17. Interleukin-1 Acts via the JNK-2 Signaling Pathway to Induce Aggrecan Degradation by Human Chondrocytes.

    Science.gov (United States)

    Ismail, Heba M; Yamamoto, Kazuhiro; Vincent, Tonia L; Nagase, Hideaki; Troeberg, Linda; Saklatvala, Jeremy

    2015-07-01

    Aggrecan enables articular cartilage to bear load and resist compression. Aggrecan loss occurs early in osteoarthritis and rheumatoid arthritis and can be induced by inflammatory cytokines such as interleukin-1 (IL-1). IL-1 induces cleavage of specific aggrecans characteristic of the ADAMTS proteinases. The aim of this study was to identify the intracellular signaling pathways by which IL-1 causes aggrecan degradation by human chondrocytes and to investigate how aggrecanase activity is controlled by chondrocytes. We developed a cell-based assay combining small interfering RNA (siRNA)-induced knockdown with aggrecan degradation assays. Human articular chondrocytes were overlaid with bovine aggrecan after transfection with siRNAs against molecules of the IL-1 signaling pathway. After IL-1 stimulation, released aggrecan fragments were detected with AGEG and ARGS neoepitope antibodies. Aggrecanase activity and tissue inhibitor of metalloproteinases 3 levels were measured by enzyme-linked immunosorbent assay. Low-density lipoprotein receptor-related protein 1 (LRP-1) shedding was analyzed by Western blotting. ADAMTS-5 is a major aggrecanase in human chondrocytes, regulating aggrecan degradation in response to IL-1. The tumor necrosis factor receptor-associated 6 (TRAF-6)/transforming growth factor β-activated kinase 1 (TAK-1)/MKK-4 signaling axis is essential for IL-1-induced aggrecan degradation, while NF-κB is not. Of the 3 MAPKs (ERK, p38, and JNK), only JNK-2 showed a significant role in aggrecan degradation. Chondrocytes constitutively secreted aggrecanase, which was continuously endocytosed by LRP-1, keeping the extracellular level of aggrecanase low. IL-1 induced aggrecanase activity in the medium in a JNK-2-dependent manner, possibly by reducing aggrecanase endocytosis, because IL-1 caused JNK-2-dependent shedding of LRP-1. The signaling axis TRAF-6/TAK-1/MKK-4/JNK-2 mediates IL-1-induced aggrecanolysis. The level of aggrecanase is controlled by its

  18. Engineering zonal cartilage through bioprinting collagen type II hydrogel constructs with biomimetic chondrocyte density gradient.

    Science.gov (United States)

    Ren, Xiang; Wang, Fuyou; Chen, Cheng; Gong, Xiaoyuan; Yin, Li; Yang, Liu

    2016-07-20

    Cartilage tissue engineering is a promising approach for repairing and regenerating cartilage tissue. To date, attempts have been made to construct zonal cartilage that mimics the cartilaginous matrix in different zones. However, little attention has been paid to the chondrocyte density gradient within the articular cartilage. We hypothesized that the chondrocyte density gradient plays an important role in forming the zonal distribution of extracellular matrix (ECM). In this study, collagen type II hydrogel/chondrocyte constructs were fabricated using a bioprinter. Three groups were created according to the total cell seeding density in collagen type II pre-gel: Group A, 2 × 10(7) cells/mL; Group B, 1 × 10(7) cells/mL; and Group C, 0.5 × 10(7) cells/mL. Each group included two types of construct: one with a biomimetic chondrocyte density gradient and the other with a single cell density. The constructs were cultured in vitro and harvested at 0, 1, 2, and 3 weeks for cell viability testing, reverse-transcription quantitative PCR (RT-qPCR), biochemical assays, and histological analysis. We found that total ECM production was positively correlated with the total cell density in the early culture stage, that the cell density gradient distribution resulted in a gradient distribution of ECM, and that the chondrocytes' biosynthetic ability was affected by both the total cell density and the cell distribution pattern. Our results suggested that zonal engineered cartilage could be fabricated by bioprinting collagen type II hydrogel constructs with a biomimetic cell density gradient. Both the total cell density and the cell distribution pattern should be optimized to achieve synergistic biological effects.

  19. [Study on the method of two dimensional polycrylamide gel electrophoresis on rat condylar chondrocyte].

    Science.gov (United States)

    Wu, Tuo-jiang; Li, Huang; Ma, Qiao-lin; Wang, Wen-mei

    2010-08-01

    To investigate the protein profile by two dimensional polycrylamide gel electrophoresis on the rat condylar chondrocyte in vitro. The third-passage chondrocytes were harvested from the mandibular condyles of 2-day-old rats in this study. The protein profile of the rat mandibular condylar chondrocytes was examined by two dimensional polycrylamide gel electrophoresis (2-DE-PAGE). The 2-DE gel maps on different pH gradients were obtained. The result of modified coomassi blue-sliver staining and sliver staining was compared using Pdquest 7.1 image analysis software. The results showed that the good protein profile of the condylar chondrocytes was obtained by standard Bio-Rad manual. The protein was mainly in the field from pH4 to pH7. The 1203±86 protein points were examined on 2-DE gel map by modified coomassi blue-sliver staining, and 1769±97 protein points was examined by sliver staining. The silver staining map showed more distinctly but higher background than modified coomassi blue-sliver staining. The protein profile of the condylar chondrocytes enriches the proteomic database and gives evidence to further proteomic research. The 2-DE map obtained by modified coomassi blue-sliver staining is more suitable for MALDI-TOF mass identification. Supported by National Natural Science Foundation of China (Grant No. C30700963), China Postdoctoral Science Foundation(Grant No.20090461088), Jiangsu Provincial Postdoctoral Science Foundation (Grant No.0802003C) and Nanjing City's Science and Technology Foundation (Grant No.200905011).

  20. Biotechnological Chondroitin a Novel Glycosamminoglycan With Remarkable Biological Function on Human Primary Chondrocytes.

    Science.gov (United States)

    Stellavato, Antonietta; Tirino, Virginia; de Novellis, Francesca; Della Vecchia, Antonella; Cinquegrani, Fabio; De Rosa, Mario; Papaccio, Gianpaolo; Schiraldi, Chiara

    2016-09-01

    Cartilage tissue engineering, with in vitro expansion of autologus chondrocytes, is a promising technique for tissue regeneration and is a new potential strategy to prevent and/or treat cartilage damage (e.g., osteoarthritis). The aim of this study was (i) to investigate and compare the effects of new biotechnological chondroitin (BC) and a commercial extractive chondroitin sulfate (CS) on human chondrocytes in vitro culture; (ii) to evaluate the anti-inflammatory effects of the innovative BC compared to extractive CS. A chondrogenic cell population was isolated from human nasoseptal cartilage and in vitro cultures were studied through time-lapse video microscopy (TLVM), immunohistochemical staining and cytometry. In order to investigate the effect of BC and CS on phenotype maintainance, chondrogenic gene expression of aggrecan (AGN), of the transcriptor factor SOX9, of the types I and II collagen (COL1A1 and COL1A2), were quantified through transcriptional and protein evaluation at increasing cultivation time and passages. In addition to resemble the osteoarthritis-like in vitro model, chondrocytes were treated with IL-1β and the anti-inflammatory activity of BC and CS was assessed using cytokines quantification by multiplex array. BC significantly enhances cell proliferation also preserving chondrocyte phenotype increasing type II collagen expression up to 10 days of treatment and reduces inflammatory response in IL-1β treated chondrocytes respect to CS treated cells. Our results, taken together, suggest that this new BC is of foremost importance in translational medicine because it can be applied in novel scaffolds and pharmaceutical preparations aiming at cartilage pathology treatments such as the osteoarthritis. J. Cell. Biochem. 117: 2158-2169, 2016. © 2016 The Authors. Journal of Cellular Biochemistry Published by Wiley Periodicals, Inc. © 2016 The Authors. Journal of Cellular Biochemistry Published by Wiley Periodicals, Inc.

  1. Chondrocyte heterogeneity: immunohistologically defined variation of integrin expression at different sites in human fetal knees.

    Science.gov (United States)

    Salter, D M; Godolphin, J L; Gourlay, M S

    1995-04-01

    During development and at maturity different forms of cartilage vary in morphology and macromolecular content. This reflects heterogeneity of chondrocyte activity, in part involving differential interactions with the adjacent extracellular matrix via specialized cell surface receptors such as integrins. We undertook an immunohistological study on a series of human fetal knee joints to assess variation in the expression of integrins by chondrocytes and potential matrix ligands in articular, epiphyseal, growth plate, and meniscal cartilage. The results show that articular chondrocytes (beta 1+, beta 5 alpha V+, alpha 1+, alpha 2+/-, alpha 5+, weakly alpha 6+, alpha V+) differed from epiphyseal (beta 1+, beta 5 alpha V+, alpha 1+/-, alpha 2+/-, alpha 5+, alpha 6+, alpha V+) growth plate (beta 1+, beta 5 alpha V+, alpha 1-, alpha 2-, alpha 5+, alpha 6+, alpha V+), and meniscal cells (beta 1+, beta 5 alpha V+, alpha 1+, strongly alpha 2+, alpha 5+, alpha 6+, alpha V+ in expression of integrin subunits. There was no expression of beta 3, beta 4, beta 6, or alpha 3 by chondrocytes. These results differ from previous reports on the expression of integrins by adult articular cartilage, where alpha 2 and alpha 6 are not seen. Variation in distribution of matrix ligands was also seen. Fibronectin, laminin and Type VI collagen were expressed in all cartilages but there was restricted expression of tenascin, ED-A and ED-B fibronectin isoforms (articular cartilage and meniscus), and vitronectin (absent from growth plate cartilage). Regulated expression of integrins by chondrocytes, associated with changes in the pericellular matrix composition, is of potential importance in control of cartilage differentiation and function in health and disease.

  2. Nanosized fibers' effect on adult human articular chondrocytes behavior

    Energy Technology Data Exchange (ETDEWEB)

    Stenhamre, Hanna [Biopolymer Technology, Department of Chemical and Biological Engineering, Chalmers University of Technology, Gothenburg (Sweden); Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska University Hospital, Gothenburg (Sweden); Thorvaldsson, Anna, E-mail: anna.thorvaldsson@swerea.se [Biopolymer Technology, Department of Chemical and Biological Engineering, Chalmers University of Technology, Gothenburg (Sweden); Swerea IVF, Mölndal (Sweden); Enochson, Lars [Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska University Hospital, Gothenburg (Sweden); Walkenström, Pernilla [Swerea IVF, Mölndal (Sweden); Lindahl, Anders [Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska University Hospital, Gothenburg (Sweden); Brittberg, Mats [Cartilage Research Unit, University of Gothenburg, Department Orthopaedics, Kungsbacka Hospital, Kungsbacka (Sweden); Gatenholm, Paul [Biopolymer Technology, Department of Chemical and Biological Engineering, Chalmers University of Technology, Gothenburg (Sweden)

    2013-04-01

    Tissue engineering with chondrogenic cell based therapies is an expanding field with the intention of treating cartilage defects. It has been suggested that scaffolds used in cartilage tissue engineering influence cellular behavior and thus the long-term clinical outcome. The objective of this study was to assess whether chondrocyte attachment, proliferation and post-expansion re-differentiation could be influenced by the size of the fibers presented to the cells in a scaffold. Polylactic acid (PLA) scaffolds with different fiber morphologies were produced, i.e. microfiber (MS) scaffolds as well as nanofiber-coated microfiber scaffold (NMS). Adult human articular chondrocytes were cultured in the scaffolds in vitro up to 28 days, and the resulting constructs were assessed histologically, immunohistochemically, and biochemically. Attachment of cells and serum proteins to the scaffolds was affected by the architecture. The results point toward nano-patterning onto the microfibers influencing proliferation of the chondrocytes, and the overall 3D environment having a greater influence on the re-differentiation. In the efforts of finding the optimal scaffold for cartilage tissue engineering, studies as the current contribute to the knowledge of how to affect and control chondrocytes behavior. - Highlights: ► Chondrocyte behavior in nanofiber-coated microfiber versus microfiber scaffolds ► High porosity (> 90%) and large pore sizes (a few hundred μm) of nanofibrous scaffolds ► Proliferation enhanced by presence of nanofibers ► Differentiation not significantly affected ► Cell attachment improved in presence of both nanofibers and serum.

  3. IKKα/CHUK regulates extracellular matrix remodeling independent of its kinase activity to facilitate articular chondrocyte differentiation.

    Directory of Open Access Journals (Sweden)

    Eleonora Olivotto

    Full Text Available BACKGROUND: The non-canonical NF-κB activating kinase IKKα, encoded by CHUK (conserved-helix-loop-helix-ubiquitous-kinase, has been reported to modulate pro- or anti- inflammatory responses, cellular survival and cellular differentiation. Here, we have investigated the mechanism of action of IKKα as a novel effector of human and murine chondrocyte extracellular matrix (ECM homeostasis and differentiation towards hypertrophy. METHODOLOGY/PRINCIPAL FINDINGS: IKKα expression was ablated in primary human osteoarthritic (OA chondrocytes and in immature murine articular chondrocytes (iMACs derived from IKKα(f/f:CreERT2 mice by retroviral-mediated stable shRNA transduction and Cre recombinase-dependent Lox P site recombination, respectively. MMP-10 was identified as a major target of IKKα in chondrocytes by mRNA profiling, quantitative RT-PCR analysis, immunohistochemistry and immunoblotting. ECM integrity, as assessed by type II collagen (COL2 deposition and the lack of MMP-dependent COL2 degradation products, was enhanced by IKKα ablation in mice. MMP-13 and total collagenase activities were significantly reduced, while TIMP-3 (tissue inhibitor of metalloproteinase-3 protein levels were enhanced in IKKα-deficient chondrocytes. IKKα deficiency suppressed chondrocyte differentiation, as shown by the quantitative inhibition of.Alizarin red staining and the reduced expression of multiple chondrocyte differentiation effectors, including Runx2, Col10a1 and Vegfa,. Importantly, the differentiation of IKKα-deficient chondrocytes was rescued by a kinase-dead IKKα protein mutant. CONCLUSIONS/SIGNIFICANCE: IKKα acts independent of its kinase activity to help drive chondrocyte differentiation towards a hypertrophic-like state. IKKα positively modulates ECM remodeling via multiple downstream targets (including MMP-10 and TIMP-3 at the mRNA and post-transcriptional levels, respectively to maintain maximal MMP-13 activity, which is required for ECM

  4. Fat Grafting for Facial Filling and Regeneration.

    Science.gov (United States)

    Coleman, Sydney R; Katzel, Evan B

    2015-07-01

    Plastic surgeons have come to realize that fat grafting can rejuvenate an aging face by restoring or creating fullness. However, fat grafting does much more than simply add volume. Grafted fat can transform or repair the tissues into which it is placed. Historically, surgeons have hesitated to embrace the rejuvenating potential of fat grafting because of poor graft take, fat necrosis, and inconsistent outcomes. This article describes fat grafting techniques and practices to assist readers in successful harvesting, processing, and placement of fat for optimal graft retention and facial esthetic outcomes. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. Design of a Sapling Branch Grafting Robot

    Directory of Open Access Journals (Sweden)

    Qun Sun

    2014-01-01

    Full Text Available The automatic sapling grafting methods and grafting robot technologies are not comprehensively studied despite the fact that they are urgently required in practice. For this reason, a sapling grafting robot is developed to implement automatic grafting for saplings. The developed grafting robot includes clipping mechanism, moving mechanism, cutting mechanism, binding mechanism, and Arduino MCU based control system, which is capable of clipping, moving, positioning, cutting, grafting, and binding saplings. Experiments show that the stock cutting efficiency is 98.4%, the scion cutting efficiency is 98.9%, the grafting efficiency is 87.3%, and the binding efficiency is 68.9%.

  6. Behaviour of oceanic 137Cs following the Fukushima Daiichi Nuclear Power Plant accident for four years simulated numerically by a regional ocean model

    Science.gov (United States)

    Tsumune, D.; Tsubono, T.; Aoyama, M.; Misumi, K.; Tateda, Y.

    2015-12-01

    A series of accidents at the Fukushima Dai-ichi Nuclear Power Plant (1F NPP) following the earthquake and tsunami of 11 March 2011 resulted in the release of radioactive materials to the ocean by two major pathways, direct release from the accident site and atmospheric deposition.We reconstructed spatiotemporal variability of 137Cs activity in the regional ocean for four years by numerical model, such as a regional scale and the North Pacific scale oceanic dispersion models, an atmospheric transport model, a sediment transport model, a dynamic biological compartment model for marine biota and river runoff model. Direct release rate of 137Cs were estimated for four years after the accident by comparing simulated results and observed activities very close to the site. The estimated total amounts of directly release was 3.6±0.7 PBq. Directly release rate of 137Cs decreased exponentially with time by the end of December 2012 and then, was almost constant. Decrease rate were quite small after 2013. The daily release rate of 137Cs was estimated to be the order of magnitude of 1010 Bq/day by the end of March 2015. The activity of directly released 137Cs was detectable only in the coastal zone after December 2012. Simulated 137Cs activities attributable to direct release were in good agreement with observed activities, a result that implies the estimated direct release rate was reasonable. There is no observed data of 137Cs activity in the ocean from 11 to 21 March 2011. Observed data of marine biota should reflect the history of 137Cs activity in this early period. We reconstructed the history of 137Cs activity in this early period by considering atmospheric deposition, river input, rain water runoff from the 1F NPP site. The comparisons between simulated 137Cs activity of marine biota by a dynamic biological compartment and observed data also suggest that simulated 137Cs activity attributable to atmospheric deposition was underestimated in this early period. The

  7. Assessing the fate of antibiotic contaminants in metal contaminated soils four years after cessation of long-term waste water irrigation

    International Nuclear Information System (INIS)

    Tamtam, Fatima; Oort, Folkert van; Le Bot, Barbara; Dinh, Tuc; Mompelat, Sophie; Chevreuil, Marc; Lamy, Isabelle; Thiry, Medard

    2011-01-01

    Spreading of urban wastewater on agricultural land may lead to concomitant input of organic and inorganic pollutants. Such multiple pollution sites offer unique opportunities to study the fate of both heavy metals and pharmaceuticals. We examined the occurrence and fate of selected antibiotics in sandy-textured soils, sampled four years after cessation of 100 years irrigation with urban wastewater from the Paris agglomeration. Previous studies on heavy metal contamination of these soils guided our sampling strategy. Six antibiotics were studied, including quinolones, with a strong affinity for organic and mineral soil components, and sulfonamides, a group of more mobile molecules. Bulk samples were collected from surface horizons in different irrigation fields, but also in subsurface horizons in two selected profiles. In surface horizons, three quinolones (oxolinic acid, nalidixic acid, and flumequine) were present in eight samples out of nine. Their contents varied spatially, but were well-correlated one to another. Their distributions showed great similarities regarding spatial distribution of total organic carbon and heavy metal contents, consistent with a common origin by wastewater irrigation. Highest concentrations were observed for sampling sites close to irrigation water outlets, reaching 22 μg kg -1 for nalidixic acid. Within soil profiles, the two antibiotic groups demonstrated an opposite behavior: quinolones, found only in surface horizons; sulfamethoxazole, detected in clay-rich subsurface horizons, concomitant with Zn accumulation. Such distribution patterns are consistent with chemical adsorption properties of the two antibiotic groups: immobilization of quinolones in the surface horizons ascribed to strong affinity for organic matter (OM), migration of sulfamethoxazole due to a lower affinity for OM and its interception and retention in electronegative charged clay-rich horizons. Our work suggests that antibiotics may represent a durable

  8. Hydrophilic/hydrophobic character of grafted cellulose

    Energy Technology Data Exchange (ETDEWEB)

    Takacs, E., E-mail: takacs@iki.kfki.h [Institute of Isotopes, Hungarian Academy of Sciences, Budapest (Hungary); Wojnarovits, L. [Institute of Isotopes, Hungarian Academy of Sciences, Budapest (Hungary); Borsa, J. [Budapest University of Technology and Economics (Hungary); Racz, I. [Bay Zoltan Institute for Materials Science and Technology, Budapest (Hungary)

    2010-04-15

    Vinyl monomers with long paraffin chains were grafted onto two kinds of cellulose (cotton and cotton linter) by direct irradiation grafting technique. The effect of dose, monomer structure and concentration, as well as homopolymer suppressor (styrene) concentration on the grafting yield was studied and the optimal grafting conditions were established. Grafting decreased the swelling of the samples in water and increased their polymer compatibility in polypropylene matrix.

  9. The prevalence of Giardia intestinalis and Entamoeba histolytica/dispar in Van Regional Training and Research Hospital: A four-year monitoring

    Directory of Open Access Journals (Sweden)

    Yasemin Bayram

    2013-03-01

    Full Text Available Objective: The aim of this retrospective study was to determinethe frequency rates of Giardia and E.histolytica/E.dispar and their distribution by years as well as agegroups and gender distribution during a four-year period.Materials and methods: A total of 9911 stool samplessent to our laboratory between January 2008 and December2011 were tested for parasites. Native-Lugol and formolethyl acetate sedimentation methods were utilized formicroscopic identification of Entamoeba sp. and Giardiatrophozoites and cysts in fresh stool samples. Additionally,trichrome staining was performed in stool sampleswhere the distinctive diagnosis could not be confirmed.Results: From a total of 9911 stool samples analyzedduring the study, 4.7% were positive for Giardia and 6.2%were positive for Entemoeba histolytica/ Entemoeba dispar.Of Giardia-positive patients 57% were male and 43%female. Similarly, 56% of Entemoeba histolytica/ Entemoebadispar positive patients were male and 44% werefemale. Both parasites’ higher frequency rates seen inmale groups were found statistically significant (p<0.01.Conclusion: It is seen that intestinal protozoon infectionsare still present as an important public health problemin our region. In order to prevent this problem, personalhygiene and sanitation rules education for community aswell as infrastructure improvements are necessary.Key words: Giardia, Entemoeba histolytica/ Entemoeba dispar, prevalence

  10. Bone Marrow Examination in Cases of New-onset Pancytopenia: A Four-year Study from a Medical College in the Rural Hilly Setting of Western Himalayas, India

    Directory of Open Access Journals (Sweden)

    Rashmi Kaul Raina

    2016-05-01

    Full Text Available New-onset pancytopenia is a common diagnostic challenge. Pancytopenia is an indication for bone marrow examination. The present study has been carried out to determine the frequencies of various etiologies of pancytopenia based on bone marrow morphology in a defined geographical location. All cases of new-onset pancytopenia, diagnosed on peripheral smear and seen over a four-year period from January 2012 to December 2015 in the department of pathology, were analysed. Patients lacking representative bone marrow in the aspirate or receiving chemotherapy were excluded. Out of 69 cases, 29 were males and 40 were females. Most of the patients were in the age group of 19-60 years (52.2%. Nineteen (26.1% of them were less than 18 years old. The three major causes of pancytopenia were: megaloblastic anemia (hypercellular marrow with megaloblastic erythropoiesis in 25 (36.2% cases, hypercellular marrow with dimorphic erythropoiesis in 13 (18.8% cases, and haematological malignancies in 12 (17.4% cases of the study. Bone marrow examination along with laboratory evaluation helps to establish specific diagnosis in cases of new-onset pancytopenia.

  11. The diet, physical activity and accommodation of patients with quiescent pulmonary tuberculosis in a poor South Indian community. A four-year follow-up study.

    Science.gov (United States)

    Ramakrishnan, C V; Rajendran, K; Mohan, K; Fox, W; Radhakrishna, S

    1966-01-01

    A previous report from the Tuberculosis Chemotherapy Centre, Madras, has shown that, if standard chemotherapy is given for one year, the response of patients treated at home in very poor environmental circumstances is nearly as good as that of those treated in sanatorium under much more favourable conditions. This paper reports on a four-year follow-up of all the patients whose disease was bacteriologically quiescent at the end of the year's treatment. During this period, all the patients were managed on a domiciliary basis; about a quarter of them received chemotherapy with isoniazid alone for two years, another quarter received the drug for one year and the rest received no specific chemotherapy. Despite adverse environmental factors (poor diet; long hours of work often involving strenuous physical activity; overcrowded living conditions; and, for the sanatorium patients, the stresses of returning suddenly to the unfavourable home environment), the great majority of patients in both series maintained quiescent disease throughout the follow-up period. Furthermore, the few patients whose disease relapsed bacteriologically were at no special dietary disadvantage in comparison with those who maintained quiescent disease throughout, nor did they show any appreciable differences in occupation, physical activity or living accommodation. These findings, together with the earlier ones, indicate that, despite adverse environmental circumstances, standard chemotherapy for an adequate period of time is sufficient in the great majority of patients for the attainment of bacteriological quiescence and its maintenance thereafter.

  12. Development of four-year fuel cycle based on the advanced fuel assembly with uranium-gadolinium fuel and its implementation to the operating WWER-440 units

    International Nuclear Information System (INIS)

    Lunin, G.; Novikov, A.; Pavlov, V.; Pavlovichev, P.; Filimonov, P.

    2000-01-01

    Over the past few years in Russia the investigations aimed at the increase of the reliability, safety and efficiency of operation of the WWER-1000 reactors as well as of its competitiveness in the world market were carried out. In the frame of these investigations the four-year fuel cycle, based on advanced fuel assemblies with zirconium alloy spacer grids and guide tubes and with fuel pellet having a reduced diameter of the central hole (1,5 mm), has been developed. For the compensation of a part of excess reactivity, Gd 2 O 3 integrated burnable absorbers are used. CPS absorbing rods contain a combine absorber (B 4 C + Dy 2 O 3 *TiO 2 ). A part of depleted fuel is located on the core periphery. The algorithms controlling the reactor power and power distribution have been updated. For checking of the solutions adopted and for verification of code package developed at the RRC 'Kurchatov Institute' the wide-scale experimental operation of advanced FA and its individual components is carried out. (Authors)

  13. Enhancing psychosocial and spiritual palliative care: Four-year results of the program of comprehensive care for people with advanced illnesses and their families in Spain.

    Science.gov (United States)

    Gómez-Batiste, Xavier; Mateo-Ortega, Dolors; Lasmarías, Cristina; Novellas, Anna; Espinosa, Jose; Beas, Elba; Ela, Sara; Barbero, Javier

    2017-02-01

    We aimed to describe the overall quantitative and qualitative results of a "La Caixa" Foundation and World Health Organization Collaborating Center Program entitled "Comprehensive Care for Patients with Advanced Illnesses and their Families" after four years of experience. Qualitative and quantitative methods were employed to assess the program. Quasiexperimental, prospective, multicenter, single-group, and pretest/posttest methods were utilized to assess the quantitative data. The effectiveness of psychosocial interventions was assessed at baseline (visit 1) and after four follow-up visits. The following dimensions were assessed: mood state, discomfort, anxiety, degree of adjustment or adaptation to disease, and suffering. We also assessed the four dimensions of the spiritual pain scale: faith or spiritual beliefs, valuable faith or spiritual beliefs, meaning in life, and peace of mind/forgiveness. Qualitative analyses were performed via surveys to evaluate stakeholder satisfaction. We built 29 psychosocial support teams involving 133 professionals-mainly psychologists and social workers. During the study period, 8,964 patients and 11,810 family members attended. Significant improvements were observed in the psychosocial and spiritual dimensions assessed. Patients, family members, and stakeholders all showed high levels of satisfaction. This model of psychosocial care could serve as an example for other countries that wish to improve psychosocial and spiritual support. Our results confirm that specific psychosocial interventions delivered by well-trained experts can help to ease suffering and discomfort in end-of-life and palliative care patients, particularly those with high levels of pain or emotional distress.

  14. Long-term leaching behavior of simulated Savannah River Plant waste glass: Part 1, MCC-1 leachability results, four-year leaching data

    International Nuclear Information System (INIS)

    Wicks, G.G.; Stone, J.A.; Chandler, G.T.; Williams, S.

    1986-08-01

    Long-term leaching data were obtained on SRP 131/TDS waste glass using MCC-1 or slightly modified MCC-1 standard leaching tests. Experiments were conducted out to four years at 40 0 C and 3-1/2 years at 90 0 C. These experiments have produced the longest standardized leaching data currently available in the waste management community. Long-term leaching data provide important input to modeling of waste glass behavior and ultimate prediction of waste glass performance. In this study, the leaching behavior of SRP waste glass was found to be excellent; leachates based on a variety of elements were not only very low, but also improved with increasing time. In addition to these data, results are also reported from another independent Savannah River study. Leaching behavior at 40 0 C and 90 0 C was assessed not only for a similar SRP 131 waste glass composition, but also for extreme waste glass compositions involving high-iron and high-aluminum waste. In addition, these experiments were performed using not only a standard deionized water leachant, but also simplified brine and silicate groundwater simulations. These two large data bases will be summarized and correlated along with some of the more interesting results recently reported in another study, a two-year leaching program performed on a similar SRP waste glass composition at Battelle Pacific Northwest Laboratories

  15. Impact on practice of a British Association for Sexual Health and HIV Sexually Transmitted Infections Foundation (STIF) course: an audit of the first four years in Ireland.

    Science.gov (United States)

    Calamai, A; Howard, R; Kelly, R; Lambert, J

    2013-02-01

    In order to investigate the overall impact of the British Association for Sexual Health and HIV (BASHH) Sexually Transmitted Infections Foundation (STIF) course taught in Ireland since 2007, attendees were sent two questionnaires to investigate the overall impact of the course, its effect on clinical practice and the need for further education. Response rate was 19.4%. The majority found the course beneficial and that it did cover their practice needs (96.4%), with 83.6% saying that their confidence and technique in sexual history taking had improved. There was a 3.7% increase in the provision of HIV testing from precourse levels, although only 80% did so routinely; a 12.7% increase in syphilis testing; a 5.4% increase in testing for Chlamydia and a 12.7% increase for gonorrhoea. Some confusion seems to persist in relation to sexually transmitted infection (STI) risk factors. The second questionnaire tested STI knowledge. Most respondents scored well (average 81% correct answers); however, respondents who attended four years previously scored, on average, 7% worse than the others, suggesting the need for a periodic update in the area of STI education.

  16. Seasonal variations and sources of atmospheric polycyclic aromatic hydrocarbons and organochlorine compounds in a high-altitude city: Evidence from four-year observations

    Energy Technology Data Exchange (ETDEWEB)

    Gong, Ping; Wang, Xiaoping; Sheng, Jiujiang; Wang, Hailong; Yuan, Xiaohua; He, Yuanqing; Qian, Yun; Yao, Tandong

    2018-02-01

    Lijiang is a high-altitude city located on the eastern fringe of the Tibetan Plateau, with complex seasonal atmospheric circulations (i.e. westerly wind, Indian Monsoon, and East Asia Monsoon). Very few previous studies have focused on seasonal variations and sources of organic pollutants in Lijiang. In this study, a four-year air campaign from June 2009 to July 2013 was conducted to investigate the temporal trends and the sources of polycyclic aromatic hydrocarbons (PAHs) and organochlorine compounds [including organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs)]. The atmospheric PAH concentrations in winter are 2-3 times of those in summer, probably because of the combined result of enhanced local emission and long-range atmospheric transport (LRAT) during winter. Traffic pollution was the primary local source of PAHs, while biomass burning is the dominant LRAT source. Different from PAHs, OCPs and PCBs mainly underwent LRAT to reach Lijiang. The peak concentrations of most of OCPs occurred in pre-monsoon season and winter, which are carried by air masses from Myanmar and India through westerly winds. As compared with other sites of the Tibetan Plateau, without the direct barrier of the Himalaya, Lijiang is easily contaminated by the incursion of polluted air masses.

  17. In-situ infiltration performance of different permeable pavements in a employee used parking lot--A four-year study.

    Science.gov (United States)

    Kumar, Kuldip; Kozak, Joseph; Hundal, Lakhwinder; Cox, Albert; Zhang, Heng; Granato, Thomas

    2016-02-01

    Permeable pavements are being adopted as a green solution in many parts of the world to manage urban stormwater quantity and quality. This paper reports on the measured in-situ infiltration performance over a four-year period since construction and use of three permeable parking sections (permeable pavers, permeable concrete and permeable asphalt) of an employee car parking lot. There was only a marginal decline in infiltration rates of all three pavements after one year of use. However, between years two to four, the infiltration rates declined significantly due to clogging of pores either by dry deposition of particles and/or shear stress of vehicles driving and degrading the permeable surfaces; during the last two years, a greater decline was also observed in driving areas of the parking lots compared to parking slots, where minimal wear and tear are expected. Maintenance strategies were employed to reclaim some of the lost infiltration rate of the permeable pavements to limited success. Despite this decline, the infiltration rates were still four to five times higher than average rainstorm intensity in the region. Thus, these permeable pavement parking lots may have significant ecological importance due to their ability to infiltrate rainwater quickly, reduce the runoff in the catchment area, and also dampen runoff peak flows that could otherwise enter the collection system for treatment in a combined sewer area. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. A hacker's guide to catching a debris flow: Lessons learned from four years of chasing mud in Colorado and southern California

    Science.gov (United States)

    Kean, J. W.; McCoy, S. W.; Staley, D. M.; Coe, J.; Leeper, R.; Tucker, G. E.

    2012-12-01

    Direct measurements of natural debris flows provide valuable insights into debris-flow processes and hazards. Yet debris flows are difficult to "catch" because they live in rugged terrain, appear infrequently, and have an appetite for destroying monitoring equipment. We present an overview of some successful (and failed) techniques we have used over the past four years to obtain direct measurements of 40+ debris flows in Colorado and southern California. Following the "MacGyver" theme of the session, we focus on the improvised equipment and methods we use in our hunt for quality data. These include an inexpensive erosion sensor to measure rates of debris-flow entrainment, a custom load cell enclosure for measuring debris-flow normal force, tracer rocks implanted with passive integrated transponders, basic pressure transducers to measure debris-flow timing, and standard digital cameras adapted to obtain high-resolution (1936 x 1288 pixels) video footage of debris flows. These techniques are also suitable for catching data on elusive flash floods. In addition, we also share some practical solutions to the logistical problems associated with installing monitoring equipment in rugged debris-flow terrain, such as suspension of non-contact stage gages high above channels.

  19. Air Pump-Assisted Graft Centration, Graft Edge Unfolding, and Graft Uncreasing in Young Donor Graft Pre-Descemet Endothelial Keratoplasty.

    Science.gov (United States)

    Jacob, Soosan; Narasimhan, Smita; Agarwal, Amar; Agarwal, Athiya; A I, Saijimol

    2017-08-01

    To assess an air pump-assisted technique for graft centration, graft edge unfolding, and graft uncreasing while performing pre-Descemet endothelial keratoplasty (PDEK) using young donor grafts. Continuous pressurized air infusion was used for graft centration, graft edge unfolding, and graft unwrinkling. Ten eyes of 10 patients underwent PDEK with donors aged below 40 years. In all eyes, the donor scrolled into tight scrolls. In all cases, the air pump-assisted technique was effective in positioning and centering the graft accurately and in straightening infolded graft edges and smoothing out graft creases and wrinkles. Endothelial cell loss was 38.6%. Postoperative best-corrected visual acuity at 6 months was 0.66 ± 0.25 in decimal equivalent. Continuous pressurized air infusion acted as a third hand providing a continuous pressure head that supported the graft and prevented graft dislocation as well as anterior chamber collapse during intraocular maneuvering. Adequate maneuvering space was available in all cases, and bleeding, if any, was tamponaded successfully in all cases. Although very young donor grafts may be used for PDEK, they are difficult to center and unroll completely before floating against host stroma. An air pump-assisted technique using continuous pressurized air infusion allows successful final graft positioning even with very young donor corneas. It thus makes surgery easier as several key steps are made easier to handle. It additionally helps in tamponading hemorrhage during peripheral iridectomy, increasing surgical space, preventing fluctuations in the anterior chamber depth, and promoting graft adherence.

  20. FAS grafted superhydrophobic ceramic membrane

    Energy Technology Data Exchange (ETDEWEB)

    Lu Jun [School of Material Science and Engineering, Jingdezhen Ceramic Institute, 333001 Jingdezhen (China); Key Laboratory of Inorganic Coating Materials, Shanghai Institute of Ceramics, CAS, 1295 DingXi Road, Shanghai 200050 (China); Yu Yun, E-mail: yunyush@mail.sic.ac.cn [Key Laboratory of Inorganic Coating Materials, Shanghai Institute of Ceramics, CAS, 1295 DingXi Road, Shanghai 200050 (China); Zhou Jianer [School of Material Science and Engineering, Jingdezhen Ceramic Institute, 333001 Jingdezhen (China); Song Lixin; Hu Xingfang [Key Laboratory of Inorganic Coating Materials, Shanghai Institute of Ceramics, CAS, 1295 DingXi Road, Shanghai 200050 (China); Larbot, Andre [Institut Europeen des Membranes, UMR 5635-CNRS, ENSCM, UMII, 1919 Route de Mende 34293, Montpellier Cedex 5 (France)

    2009-08-30

    The hydrophobic properties of {gamma}-Al{sub 2}O{sub 3} membrane have been obtained by grafting fluoroalkylsilane (FAS) on the surface of the membrane. The following grafting parameters were studied: the eroding time of the original membrane, the grafting time, the concentration of FAS solution and the multiplicity of grafting. Hydrophobicity of the membranes was characterized by contact angle (CA) measurement. The thermogravimetric analysis (TGA) was used to investigate the weight loss process (25-800 deg. C) of the fluoroalkylsilane grafted on Al{sub 2}O{sub 3} powders under different grafting conditions. The morphologies of the membranes modified under different parameters were examined by field emission scanning electron microscopy (FE-SEM) and the surface roughness (Ra) was measured using white light interferometers. A needle-like structure was observed on the membrane surface after modification, which causes the change of Ra. On the results above, we speculated a model to describe the reaction between FAS and {gamma}-Al{sub 2}O{sub 3} membrane surface as well as the formed surface morphology.

  1. FAS grafted superhydrophobic ceramic membrane

    Science.gov (United States)

    Lu, Jun; Yu, Yun; Zhou, Jianer; Song, Lixin; Hu, Xingfang; Larbot, Andre

    2009-08-01

    The hydrophobic properties of γ-Al 2O 3 membrane have been obtained by grafting fluoroalkylsilane (FAS) on the surface of the membrane. The following grafting parameters were studied: the eroding time of the original membrane, the grafting time, the concentration of FAS solution and the multiplicity of grafting. Hydrophobicity of the membranes was characterized by contact angle (CA) measurement. The thermogravimetric analysis (TGA) was used to investigate the weight loss process (25-800 °C) of the fluoroalkylsilane grafted on Al 2O 3 powders under different grafting conditions. The morphologies of the membranes modified under different parameters were examined by field emission scanning electron microscopy (FE-SEM) and the surface roughness (Ra) was measured using white light interferometers. A needle-like structure was observed on the membrane surface after modification, which causes the change of Ra. On the results above, we speculated a model to describe the reaction between FAS and γ-Al 2O 3 membrane surface as well as the formed surface morphology.

  2. Interventions in Infrainguinal Bypass Grafts

    International Nuclear Information System (INIS)

    Mueller-Huelsbeck, S.; Order, B.-M.; Jahnke, T.

    2006-01-01

    The interventional radiologist plays an important role in the detection and prevention of infrainguinal bypass failure. Early detection and evaluation of flow-limiting lesions effectively preserve graft (venous bypass and polyester or expanded polytetrafluoroethylene bypass) patency by identifying stenoses before occlusion occurs. Delay in treatment of the at-risk graft may result in graft failure and a reduced chance of successful revascularization. For this reason, surveillance protocols form an important part of follow-up after infrainguinal bypass surgery. As well as having an understanding of the application of imaging techniques including ultrasound, MR angiography, CT angiography and digital subtraction angiography, the interventional radiologist should have detailed knowledge of the minimally invasive therapeutic options. Percutaneous transluminal angioplasty (PTA), or alternatively cutting balloon angioplasty, is the interventional treatment of choice in prevention of graft failure and occlusion. Further alternatives include metallic stent placement, fibrinolysis, and mechanical thrombectomy. Primary assisted patency rates following PTA can be up to 65% at 5 years. When the endovascular approach is unsuccessful, these therapeutic options are complemented by surgical procedures including vein patch revision, jump grafting, or placement of a new graft

  3. Constitutive activation of MEK1 in chondrocytes causes Stat1-independent achondroplasia-like dwarfism and rescues the Fgfr3-deficient mouse phenotype

    Science.gov (United States)

    Murakami, Shunichi; Balmes, Gener; McKinney, Sandra; Zhang, Zhaoping; Givol, David; de Crombrugghe, Benoit

    2004-01-01

    We generated transgenic mice that express a constitutively active mutant of MEK1 in chondrocytes. These mice showed a dwarf phenotype similar to achondroplasia, the most common human dwarfism, caused by activating mutations in FGFR3. These mice displayed incomplete hypertrophy of chondrocytes in the growth plates and a general delay in endochondral ossification, whereas chondrocyte proliferation was unaffected. Immunohistochemical analysis of the cranial base in transgenic embryos showed reduced staining for collagen type X and persistent expression of Sox9 in chondrocytes. These observations indicate that the MAPK pathway inhibits hypertrophic differentiation of chondrocytes and negatively regulates bone growth without inhibiting chondrocyte proliferation. Expression of a constitutively active mutant of MEK1 in chondrocytes of Fgfr3-deficient mice inhibited skeletal overgrowth, strongly suggesting that regulation of bone growth by FGFR3 is mediated at least in part by the MAPK pathway. Although loss of Stat1 restored the reduced chondrocyte proliferation in mice expressing an achondroplasia mutant of Fgfr3, it did not rescue the reduced hypertrophic zone, the delay in formation of secondary ossification centers, and the achondroplasia-like phenotype. These observations suggest a model in which Fgfr3 signaling inhibits bone growth by inhibiting chondrocyte differentiation through the MAPK pathway and by inhibiting chondrocyte proliferation through Stat1. PMID:14871928

  4. Photobiostimulation on chondrocytes proliferation in different concentration of fetal bovine serum under low-level laser irradiation

    Science.gov (United States)

    Zheng, Liqin; Wang, Yuhua; Qiu, Caimin; Chen, Jianlin; Yang, Hongqin; Zhang, Yanding; Xie, Shusen

    2015-03-01

    The aim of this in vitro study was to evaluate the influence of low-level laser irradiation (LLLI) on the chondrocytes proliferation cultured in different concentration of fetal bovine serum (FBS) using 658 nm, 785 nm and 830 nm diode lasers. The role of energy density (10-70 mJ·cm-2) on chondrocytes proliferation following irradiation with 658 nm laser for 2 days was firstly investigated to find out the best laser energy density. Then the effect of LLLI on the proliferation of chondrocytes cultured with fetal bovine serum at 0%, 2%, 5% and 10% was also evaluated. The results showed that there was no or little photobiostimulation on the proliferation of chondrocytes cultured with 0% FBS and 10% FBS; the cell proliferation at 2% and 5% FBS was significantly modulated by LLLI.

  5. Incorporation of hyaluronic acid into collagen scaffolds for the control of chondrocyte-mediated contraction and chondrogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Tang Shunqing [Department of Biomedical Engineering, Jinan University, Guangzhou 510632 (China); Spector, Myron [Tissue Engineering, VA Boston Healthcare System, Boston, MA 02130 (United States)

    2007-09-15

    Hyaluronic acid (HA), a principal matrix molecule in many tissues, is present in high amounts in articular cartilage. HA contributes in unique ways to the physical behavior of the tissue, and has been shown to have beneficial effects on chondrocyte activity. The goal of this study was to incorporate graduated amounts of HA into type I collagen scaffolds for the control of chondrocyte-mediated contraction and chondrogenesis in vitro. The results demonstrated that the amount of contraction of HA/collagen scaffolds by adult canine articular chondrocytes increased with the HA content of the scaffolds. The greatest amount of chondrogenesis after two weeks was found in the scaffolds which had undergone the most contraction. HA can play a useful role in adjusting the mechanical behavior of tissue engineering scaffolds and chondrogenesis in chondrocyte-seeded scaffolds.

  6. In vitro cell quality of articular chondrocytes assigned for autologous implantation in dependence of specific patient characteristics

    DEFF Research Database (Denmark)

    Pestka, Jan M; Schmal, Hagen; Salzmann, Gian

    2011-01-01

    OBJECTIVE: Autologous chondrocyte implantation (ACI) is a well-established therapeutic option for the treatment of cartilage defects of the knee joint. Since information concerning the cellular aspects of ACI is still limited, the aim of the present study was to investigate relevant differences...... between chondrocyte quality after in vitro cultivation and possible correlations with patient-specific factors. DESIGN: Cell quality of 252 consecutive ACI patients was assessed after chondrocyte in vitro expansion by determination of the expression of cartilage relevant surface marker CD44 and cartilage......, aggrecan or collagen type II nor cell density or viability after proliferation seemed to correlate with the grade of joint degeneration, defect aetiology or patient gender. However, chondrocytes harvested from the knee joints of patients at less than 20 years of age showed significantly higher expression...

  7. Sporting Activity Is Reduced 11 Years After First-Generation Autologous Chondrocyte Implantation in the Knee Joint

    DEFF Research Database (Denmark)

    Erdle, Benjamin; Herrmann, Simon; Porichis, Stella

    2017-01-01

    BACKGROUND: Little is known about long-term sporting activity after periosteal autologous chondrocyte implantation (ACI-P) and its correlation to clinical, morphological, and ultrastructural cartilage characteristics on magnetic resonance imaging (MRI). PURPOSE: To evaluate long-term sporting...

  8. SOX9 governs differentiation stage-specific gene expression in growth plate chondrocytes via direct concomitant transactivation and repression.

    Directory of Open Access Journals (Sweden)

    Victor Y L Leung

    2011-11-01

    Full Text Available Cartilage and endochondral bone development require SOX9 activity to regulate chondrogenesis, chondrocyte proliferation, and transition to a non-mitotic hypertrophic state. The restricted and reciprocal expression of the collagen X gene, Col10a1, in hypertrophic chondrocytes and Sox9 in immature chondrocytes epitomise the precise spatiotemporal control of gene expression as chondrocytes progress through phases of differentiation, but how this is achieved is not clear. Here, we have identified a regulatory element upstream of Col10a1 that enhances its expression in hypertrophic chondrocytes in vivo. In immature chondrocytes, where Col10a1 is not expressed, SOX9 interacts with a conserved sequence within this element that is analogous to that within the intronic enhancer of the collagen II gene Col2a1, the known transactivation target of SOX9. By analysing a series of Col10a1 reporter genes in transgenic mice, we show that the SOX9 binding consensus in this element is required to repress expression of the transgene in non-hypertrophic chondrocytes. Forced ectopic Sox9 expression in hypertrophic chondrocytes in vitro and in mice resulted in down-regulation of Col10a1. Mutation of a binding consensus motif for GLI transcription factors, which are the effectors of Indian hedgehog signaling, close to the SOX9 site in the Col10a1 regulatory element, also derepressed transgene expression in non-hypertrophic chondrocytes. GLI2 and GLI3 bound to the Col10a1 regulatory element but not to the enhancer of Col2a1. In addition to Col10a1, paired SOX9-GLI binding motifs are present in the conserved non-coding regions of several genes that are preferentially expressed in hypertrophic chondrocytes and the occurrence of pairing is unlikely to be by chance. We propose a regulatory paradigm whereby direct concomitant positive and negative transcriptional control by SOX9 ensures differentiation phase-specific gene expression in chondrocytes. Discrimination between

  9. Radiation grafting on natural films

    International Nuclear Information System (INIS)

    Lacroix, M.; Khan, R.; Senna, M.; Sharmin, N.; Salmieri, S.; Safrany, A.

    2014-01-01

    Different methods of polymer grafting using gamma irradiation are reported in the present study for the preparation of newly functionalized biodegradable films, and some important properties related to their mechanical and barrier properties are described. Biodegradable films composed of zein and poly(vinyl alcohol) (PVA) were gamma-irradiated in presence of different ratios of acrylic acid (AAc) monomer for compatibilization purpose. Resulting grafted films (zein/PVA-g-AAc) had their puncture strength (PS=37–40 N mm −1 ) and puncture deformation (PD=6.5–9.8 mm) improved for 30% and 50% PVA in blend, with 5% AAc under 20 kGy. Methylcellulose (MC)-based films were irradiated in the presence of 2-hydroxyethyl methacrylate (HEMA) or silane, in order to determine the effect of monomer grafting on the mechanical properties of films. It was found that grafted films (MC-g-HEMA and MC-g-silane) using 35% monomer performed higher mechanical properties with PS values of 282–296 N mm −1 and PD of 5.0–5.5 mm under 10 kGy. Compatibilized polycaprolactone (PCL)/chitosan composites were developed via grafting silane in chitosan films. Resulting trilayer grafted composite film (PCL/chitosan-g-silane/PCL) presented superior tensile strength (TS=22 MPa) via possible improvement of interfacial adhesion (PCL/chitosan) when using 25% silane under 10 kGy. Finally, MC-based films containing crystalline nanocellulose (CNC) as a filling agent were prepared and irradiated in presence of trimethylolpropane trimethacrylate (TMPTMA) as a grafted plasticizer. Grafted films (MC-g-TMPTMA) presented superior mechanical properties with a TS of 47.9 MPa and a tensile modulus (TM) of 1792 MPa, possibly due to high yield formation of radicals to promote TMPTMA grafting during irradiation. The addition of CNC led to an additional improvement of the barrier properties, with a significant 25% reduction of water vapor permeability (WVP) of grafted films. - Highlights: • Irradiation of zein

  10. Lipofection of rabbit chondrocytes and long lasting expression of a lacZ reporter system in alginate beads.

    Science.gov (United States)

    Stöve, J; Fiedler, J; Huch, K; Günther, K-P; Puhl, W; Brenner, R

    2002-03-01

    Our aim was to investigate the maintenance of the transfection status of non-viral transfected chondrocytes in an alginate culture system. Chondrocytes harvested from rabbit knees were isolated by sequential digestion and cultivated in monolayer culture. At 60-70% cell density, chondrocytes were transfected with different transfection systems (FuGENE6, CaCl2, Lipofectin). A lac Z expression vector (pcDNA 3.1/Myc-His+ lacZ) was used as a reporter system. In order to improve transfection rates, hyaluronidase (4 U/ml) was used prior and during the transfection procedure. Thereafter, transfected cells were either kept in monolayer culture or embedded in alginate beads and kept in culture for up to the next 30 weeks. Transfection efficiency was maximal using FuGENE6TM/DNA at a ratio of 3:2 and hyaluronidase (4 U/ml). Transfection efficiency reached up to 40.8% (+/- 3.2%) after 36 h. In alginate beads lac Z positive cells declined to 8.5% +/- 3.3% after 4 weeks and to 4.6% +/- 3.2% after 12 weeks of culturing. After 30 weeks 3% of chondrocytes still expressed lac Z. In contrast, during culturing in monolayer, no lac Z expression was detectable after 4 weeks. Differentiation status of the chondrocytes was confirmed by histology and immunohistochemistry methods. After successful gene transfer to rabbit chondrocytes the alginate system made it possible to culture lipofected chondrocytes phenotypically stable. Genetically engineered chondrocytes express the lac Z reporter gene over a period of at least 30 weeks. This transfection and culture system provides a promising tool to further investigate the over-expression of growth factors and enzyme inhibitors. Copyright 2002 OsteoArthritis Research Society International.

  11. The effect of dexamethasone and triiodothyronine on terminal differentiation of primary bovine chondrocytes and chondrogenically differentiated mesenchymal stem cells.

    Science.gov (United States)

    Randau, Thomas M; Schildberg, Frank A; Alini, Mauro; Wimmer, Matthias D; Haddouti, El-Mustapha; Gravius, Sascha; Ito, Keita; Stoddart, Martin J

    2013-01-01

    The newly evolved field of regenerative medicine is offering solutions in the treatment of bone or cartilage loss and deficiency. Mesenchymal stem cells, as well as articular chondrocytes, are potential cells for the generation of bone or cartilage. The natural mechanism of bone formation is that of endochondral ossification, regulated, among other factors, through the hormones dexamethasone and triiodothyronine. We investigated the effects of these hormones on articular chondrocytes and chondrogenically differentiated mesenchymal stem cells, hypothesizing that these hormones would induce terminal differentiation, with chondrocytes and differentiated stem cells being similar in their response. Using a 3D-alginate cell culture model, bovine chondrocytes and chondrogenically differentiated stem cells were cultured in presence of triiodothyronine or dexamethasone, and cell proliferation and extracellular matrix production were investigated. Collagen mRNA expression was measured by real-time PCR. Col X mRNA and alkaline phosphatase were monitored as markers of terminal differentiation, a prerequisite of endochondral ossification. The alginate culture system worked well, both for the culture of chondrocytes and for the chondrogenic differentiation of mesenchymal stem cells. Dexamethasone led to an increase in glycosaminoglycan production. Triiodothyronine increased the total collagen production only in chondrocytes, where it also induced signs of terminal differentiation, increasing both collagen X mRNA and alkaline phosphatase activity. Dexamethasone induced terminal differentiation in the differentiated stem cells. The immature articular chondrocytes used in this study seem to be able to undergo terminal differentiation, pointing to their possible role in the onset of degenerative osteoarthritis, as well as their potential for a cell source in bone tissue engineering. When chondrocyte-like cells, after their differentiation, can indeed be moved on towards terminal

  12. Reduced primary cilia length and altered Arl13b expression are associated with deregulated chondrocyte Hedgehog signaling in alkaptonuria.

    Science.gov (United States)

    Thorpe, Stephen D; Gambassi, Silvia; Thompson, Clare L; Chandrakumar, Charmilie; Santucci, Annalisa; Knight, Martin M

    2017-09-01

    Alkaptonuria (AKU) is a rare inherited disease resulting from a deficiency of the enzyme homogentisate 1,2-dioxygenase which leads to the accumulation of homogentisic acid (HGA). AKU is characterized by severe cartilage degeneration, similar to that observed in osteoarthritis. Previous studies suggest that AKU is associated with alterations in cytoskeletal organization which could modulate primary cilia structure/function. This study investigated whether AKU is associated with changes in chondrocyte primary cilia and associated Hedgehog signaling which mediates cartilage degradation in osteoarthritis. Human articular chondrocytes were obtained from healthy and AKU donors. Additionally, healthy chondrocytes were treated with HGA to replicate AKU pathology (+HGA). Diseased cells exhibited shorter cilia with length reductions of 36% and 16% in AKU and +HGA chondrocytes respectively, when compared to healthy controls. Both AKU and +HGA chondrocytes demonstrated disruption of the usual cilia length regulation by actin contractility. Furthermore, the proportion of cilia with axoneme breaks and bulbous tips was increased in AKU chondrocytes consistent with defective regulation of ciliary trafficking. Distribution of the Hedgehog-related protein Arl13b along the ciliary axoneme was altered such that its localization was increased at the distal tip in AKU and +HGA chondrocytes. These changes in cilia structure/trafficking in AKU and +HGA chondrocytes were associated with a complete