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Find video protocols related to scientific articles indexed in Pubmed.
Comparison of UVB and UVC effects on the DNA damage-response protein 53BP1 in human pancreatic cancer.
J. Cell. Biochem.
PUBLISHED: 04-23-2014
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We have previously demonstrated that ultraviolet (UV) light is effective against a variety of cancer cells expressing fluorescent proteins in vivo as well as in vitro. In the present report, we compared the DNA damage repair (DDR) response of pancreatic cancer cells after UVB or UVC irradiation. The UV-induced DNA damage repair was imaged with green fluorescent protein (GFP) fused to the DDR-related chromatin-binding protein 53BP1 in MiaPaCa-2 human pancreatic cancer cells growing in 3D Gelfoam® histoculture and in superficial tumors grown in nude mice. 53BP1-GFP forms foci during DNA damage repair. A clonogenic assay in 2D monolayer culture initially showed that UVC and UVB inhibited MiaPaCa-2 cell proliferation in a dose-dependent manner, with UVC having more efficacy. Three-dimensional Gelfoam® histocultures and confocal imaging enabled 53BP1-GFP foci to be observed within 1?h after UV irradiation, indicating the onset of DDR response. UVB-induced 53BP1-GFP focus formation was observed up to a depth of 120?µm in MiaPaCa-2 cells on Gelfoam® compared to 80?µm for UVC. UVB-induced 53BP1-GFP focus formation was observed up to a depth of 80?µm in MiaPaCa-2 cells, implanted within skin flaps in mice, at a significantly greater extent than UVC. MiaPaCa-2 cells irradiated by UVB or UVC in the skin-flap mouse model had a significant decrease in tumor growth compared to untreated controls with UVB having more efficacy than UVC. Our results demonstrate that UVB has greater tissue penetration than UVC because of its longer wavelength and has clinical potential for eradicating superficial cancer.
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Osteosarcoma cells enhance angiogenesis visualized by color-coded imaging in the in vivo Gelfoam® assay.
J. Cell. Biochem.
PUBLISHED: 02-24-2014
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We previously described a color-coded imaging model that can quantify the length of nascent blood vessels using Gelfoam® implanted in nestin-driven green fluorescent protein (ND-GFP) nude mice. In ND-GFP mice, nascent blood vessels are labeled with GFP. We report here that osteosarcoma cells promote angiogenesis in the Gelfoam® angiogenesis assay in ND-GFP mice. Gelfoam® was initially transplanted subcutaneously in the flank of transgenic ND-GFP nude mice. Seven days after transplantation of Gelfoam®, skin flaps were made and human 143B osteosarcoma cells expressing green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in cytoplasm were injected into the transplanted Gelfoam®. The control-group mice had only implanted Gelfoam®. Skin flaps were made at days 14, 21, and 28 after transplantation of the Gelfoam® to allow imaging of vascularization in the Gelfoam® using a variable-magnification small animal imaging system and confocal fluorescence microscopy. ND-GFP expressing nascent blood vessels penetrated and spread into the Gelfoam® in a time-dependent manner in both control and osteosarcoma-implanted mice. ND-GFP expressing blood vessels in the Gelfoam® of the osteosarcoma-implanted mice were associated with the cancer cells and larger and longer than in the Gelfoam®-only implanted mice (P?
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3-dimensional tissue is formed from cancer cells in vitro on Gelfoam®, but not on Matrigel™.
J. Cell. Biochem.
PUBLISHED: 01-27-2014
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Cell and tissue culture can be performed on different substrates such as on plastic, in Matrigel™, and on Gelfoam(®), a sponge matrix. Each of these substrates consists of a very different surface, ranging from hard and inflexible, a gel, and a sponge-matrix, respectively. Folkman and Moscona found that cell shape was tightly coupled to DNA synthesis and cell growth. Therefore, the flexibility of a substrate is important for cells to maintain their optimal shape. Human osteosarcoma cells, stably expressing a fusion protein of ?(v) integrin and green fluorescent protein (GFP), grew as a simple monolayer without any structure formation on the surface of a plastic dish. When the osteosarcoma cells were cultured within Matrigel™, the cancer cells formed colonies but no other structures. When the cancer cells were seeded on Gelfoam(®), the cells formed three-dimensional tissue-like structures. The behavior of 143B osteosarcoma cells on Gelfoam(®) in culture is remarkably different from those of these cells in monolayer culture or in Matrigel™. Tissue-like structures were observed only in Gelfoam(®) culture. The data in this report suggest a flexible structural substrate such as Gelfoam(®) provides a more in vivo-like culture condition than monolayer culture or Matrigel(TM) and that Matrigel(TM) does not result in actual three-dimensional culture.
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High lung-metastatic variant of human osteosarcoma cells, selected by passage of lung metastasis in nude mice, is associated with increased expression of ?(v)?(3) integrin.
Anticancer Res.
PUBLISHED: 09-12-2013
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Altered expression of ?v?3 integrin is associated with tumor progression and metastasis in several types of cancer, including metastatic osteosarcoma. In this study, we demonstrate that in vivo passaging of lung metastasis in nude mice can generate an aggressive variant of human osteosarcoma cells. Experimental metastases were established by injecting 143B human osteosarcoma cells, expressing green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in the cytoplasm, in the tail vein of nude mice. Lung metastases were harvested under fluorescence microscopy from nude mice to establish cell lines which were then injected via the tail vein of additional nude mice. This procedure was repeated for four passages in order to isolate highly metastatic variant sublines. When the parental and metastatic variants were transplanted orthotopically into the tibia of nude mice, the 143B-LM4 variant had the highest metastatic rate, approximately 18-fold higher than the parent (p<0.01). ?v?3 integrin expression was increased approximately 5.6-fold in 143B-LM4 compared to parental cells (p<0.05). Thus, serial passage of lung metastases created a highly metastatic variant of human osteosarcoma cells which had increased expression of ?v?3 integrin, suggesting that ?v?3 integrin plays an essential role in osteosarcoma metastasis. With this highly metastatic variant overexpressing ?v?3 integrin, it will now be possible to further investigate the mechanism by which ?v?3 integrin facilitates metastasis.
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Color-coded imaging of spontaneous vessel anastomosis in vivo.
Anticancer Res.
PUBLISHED: 07-31-2013
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Vessel anastomosis is important in tumor angiogenesis as well as for vascularization therapy for ischemia and other diseases. We report here the development of a color-coded imaging model that can visualize the anastomosis between blood vessels of red fluorescent protein (RFP)-expressing vessels in vascularized Gelfoam® previously transplanted into RFP transgenic mice and then re-transplanted into nestin-driven green fluorescent protein (ND-GFP) mice where nascent blood vessels express GFP. Gelfoam® was initially transplanted subcutaneously in the flank of transgenic RFP nude mice. Skin flaps were made at 14 days after transplantation of Gelfoam® to allow observation of vascularization of the Gelfoam® using confocal fluorescence imaging. The implanted Gelfoam® became highly vascularized with RFP vessels. Fourteen days after transplantation into RFP transgenic nude mice, the Gelfoam® was removed and re-transplanted into the subcutis on the flank of ND-GFP transgenic nude mice in which nascent blood vessels express GFP. Skin flaps were made and anastomosis between the GFP-expressing nascent blood vessels of ND-GFP transgenic nude mice and RFP blood vessels in the Gelfoam® was imaged 14 and 21 days after re-transplantation. The results presented in this report indicate a possible mechanism for tumor angiogenesis and suggest a new paradigm of therapeutic revascularization of ischemic organs requiring new blood vessels and in other diseases.
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Real-time imaging of ?v integrin molecular dynamics in osteosarcoma cells in vitro and in vivo.
Anticancer Res.
PUBLISHED: 07-31-2013
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?v Integrin is involved in various steps of cancer metastasis. In this report, we describe real-time imaging of ?v integrin molecular dynamics in human 143B osteosarcoma cells in vitro and in vivo. We first generated osteosarcoma cells expressing ?v integrin green fluorescent protein (GFP) by transfection of an ?v integrin GFP fusion vector (pCMV6-AC-ITGAV-GFP) into 143B cells. Confocal laser-scanning microscopy demonstrated that ?v integrin immunofluorescence staining co-localized with ?v integrin-GFP fluorescence in 143B cells. When ?v integrin-GFP-expressing 143B osteosarcoma cells were seeded on a dish coated with fibronectin, which is bound by ?v integrin, punctate ?v integrin-GFP was observed by confocal laser-scanning microscopy. When the 143B ?v integrin-GFP cells were seeded onto uncoated plastic, ?v integrin-GFP was diffuse within the cells. When ?v integrin-GFP 143B osteosarcoma cells (1×10(6)) were orthotopically transplanted into the tibia of nude mice, the cells aligned along the collagen fibers within the tumor and had punctuate expression of ?v integrin-GFP. In the orthotopic model, the invading osteosarcoma cells had punctate ?v integrin-GFP in the muscle tissue at the primary tumor margin. These results show that ?v integrin-GFP enables the imaging of the molecular dynamics of ?v integrin in osteosarcoma cells in vitro and in vivo.
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Primer dosing of S. typhimurium A1-R potentiates tumor-targeting and efficacy in immunocompetent mice.
Anticancer Res.
PUBLISHED: 06-01-2013
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We developed the tumor-targeting strain Salmonella typhimuium A1-R (A1-R) and have shown it to be active against a number of tumor types in nude mice. However, in immunocompetent mice, dosing of A1-R has to be adjusted to avoid toxicity. In the present study, we developed a strategy to maximize efficacy and minimize toxicity for A1-R tumor-targeting in immunocompetent mice implanted with the Lewis lung carcinoma. A small primer dose of A1-R was first administered (1×10(6) colony forming unit [cfu] i.v.) followed by a high dose (1×10(7) cfu i.v.) four hours later. The primer-dose strategy resulted in smaller tumors and no observable side-effects compared to treatment with high-dose-alone. The serum level of tumor necrosis factor (TNF-?) was elevated in the mice treated with primer dose compared to mice only given the high dose. Tumor vessel destruction was enhanced by primer dosing of A1-R in immuno-competent transgenic mice expressing the nestin-driven green fluorescent protein, which is selectively expressed in nascent blood vessels. The primer-dose may activate TNF-? and other cytokines in the mouse, necessary for invasion of the tumor by the bacteria, as well as enhance tumor vessel destruction, thereby allowing a subsequent therapeutic dose to be effective and safe. The results of the present study suggest effective future clinical strategies of bacterial treatment of cancer.
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Anticancer effects of marine carotenoids, fucoxanthin and its deacetylated product, fucoxanthinol, on osteosarcoma.
Int. J. Oncol.
PUBLISHED: 04-25-2013
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Survival of osteosarcoma patients hinges on prevention or treatment of recurrent and metastatic lesions. Therefore, novel chemotherapeutics for more effective treatment and prevention of this disease are required. Carotenoids are natural pigments and exhibit various biological functions. We evaluated the anti-osteosarcoma properties of several carotenoids. Among carotenoids, fucoxanthin and its metabolite fucoxanthinol, inhibited the cell viability of osteosarcoma cell lines. Fucoxanthinol induced G1 cell cycle arrest by reducing the expression of cyclin-dependent kinase 4, cyclin-dependent kinase 6 and cyclin E and apoptosis by reducing the expression of survivin, XIAP, Bcl-2 and Bcl-xL. Apoptosis was associated with activation of caspases-3, -8 and -9. In addition, fucoxanthinol inhibited the phosphorylation of phosphoinositide-dependent kinase 1 and Akt and the downstream glycogen synthase kinase 3?, resulting in downregulation of ?-catenin. Fucoxanthinol inhibited the cell migration and invasion of osteosarcoma cells. It also reduced matrix metalloproteinase-1 expression and the activator protein-1 signal. Treatment of mice inoculated with osteosarcoma cells with fucoxanthin inhibited the development of osteosarcoma in mice. Fucoxanthin and fucoxanthinol inhibit cell growth, migration and invasion and induce apoptosis of osteosarcoma cells at least in part by inhibiting Akt and activator protein-1 pathways. Our findings provide a rationale for clinical evaluation of these novel agents in osteosarcoma.
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A color-coded imaging model of the interaction of ?v integrin-GFP expressed in osteosarcoma cells and RFP expressing blood vessels in Gelfoam® vascularized in vivo.
Anticancer Res.
PUBLISHED: 04-09-2013
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The integrin family of proteins has been shown to be involved in the malignant behavior of cells. We report here development of a color-coded imaging model that can visualize the interaction between ?v integrin linked to green fluorescent protein (GFP) in osteosarcoma cells and blood vessels in Gelfoam® vascularized after implantation in red fluorescent protein (RFP) transgenic nude mice. Human 143B osteosarcoma cells expressing ?v integrin-GFP were generated by transfection with an ?v integrin-GFP vector. Gelfoam® (5×5 mm) was transplanted subcutaneously in transgenic RFP nude mice. The implanted Gelfoam® became highly vascularized with RFP vessels within 14 days. Skin flaps were made at days 7, 14, 21, 28 after transplantation of Gelfoam® for observing vascularization of the Gelfoam® using fluorescence imaging. Gelfoam® is a useful tool to observe angiogenesis in vivo. 143B cells (5 × 10(5)) expressing ?v integrin-GFP were injected into the Gelfoam® seven days after transplantation of Gelfoam®. Seven days after cancer-cell injection, cancer cells and blood vessels were observed in the Gelfoam® by color-coded confocal microscopy via the skin flap. The 143B cells expressing ?v integrin-GFP proliferated into the Gelfoam®, which contained RFP-expressing blood vessels. Strong expression of ?v integrin-GFP in 143B cells was observed near RFP vessels in the Gelfoam®. The observation of the behavior of ?v integrin-GFP and blood vessels will allow further understanding of the role of ?v integrin in cancer cells.
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Influence of the gel thickness on in vivo hyaline cartilage regeneration induced by double-network gel implanted at the bottom of a large osteochondral defect: short-term results.
BMC Musculoskelet Disord
PUBLISHED: 01-29-2013
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A double-network (DN) gel, which is composed of poly(2-acrylamido-2-methylpropanesulfonic acid) and poly(N,N-dimethyl acrylamide), can induce hyaline cartilage regeneration in vivo in a large osteochondral defect. The purpose of this study was to clarify the influence of the thickness of the implanted DN gel on the induction ability of hyaline cartilage regeneration.
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Open repair for massive rotator cuff tear with a modified transosseous-equivalent procedure: preliminary results at short-term follow-up.
J Orthop Sci
PUBLISHED: 01-24-2011
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Many surgical procedures have been reported for rotator cuff tears. We adopted the modified transosseous-equivalent procedure, also termed "surface-holding repair with transosseous sutures," and demonstrated that this procedure has a biomechanical advantage regarding the concentration of stress on the tendon stump. This study aimed to evaluate the clinical and structural outcomes of this technique, which has been demonstrated by postoperative magnetic resonance imaging (MRI) to produce high intact rates.
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Joint immobilization inhibits spontaneous hyaline cartilage regeneration induced by a novel double-network gel implantation.
J Mater Sci Mater Med
PUBLISHED: 08-26-2010
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We have recently discovered that spontaneous hyaline cartilage regeneration can be induced in an osteochondral defect in the rabbit, when we implant a novel double-network (DN) gel plug at the bottom of the defect. To clarify whether joint immobilization inhibits the spontaneous hyaline cartilage regeneration, we conducted this study with 20 rabbits. At 4 or 12 weeks after surgery, the defect in the mobile knees was filled with a sufficient volume of the hyaline cartilage tissue rich in proteoglycan and type-2 collagen, while no cartilage tissues were observed in the defect in the immobilized knees. Type-2 collagen, Aggrecan, and SOX9 mRNAs were expressed only in the mobile knees at each period. This study demonstrated that joint immobilization significantly inhibits the spontaneous hyaline cartilage regeneration induced by the DN gel implantation. This fact suggested that the mechanical environment is one of the significant factors to induce this phenomenon.
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Epidemiology of hip fractures in Okinawa, Japan.
J. Bone Miner. Metab.
PUBLISHED: 07-19-2010
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This study investigated the current incidence of hip fractures in Okinawa prefecture and compared the data with those obtained in our previous study, which was conducted using similar methods in 1987/1988. All patients, aged 50 years or older and residing in Okinawa, admitted to Okinawa hospitals in 2004 for a fresh hip fracture were identified from hospital registries. Details were obtained from the medical records and radiographs of all patients and classified according to fracture type (cervical or trochanteric), age, sex, and fracture location. Subtrochanteric fractures and pathological fractures were excluded. A total of 1,349 patients (242 men and 1,107 women) were admitted for a fresh hip fracture in 2004. Their average age was 76.9 years for men and 82.4 years for women. There were 671 cervical fractures, 654 trochanteric fractures, and 24 unclassified proximal femoral fractures. Comparing the data from 1987/1988 to those from 2004, the total number of hip fractures increased by 188%, from 469 to 1,349. The age-adjusted incidence rates per 100,000, standardized to the 2000 US population, were 75.7 and 296.1 in 1987/1988 and 123.6 and 420 in 2004 for men and women, respectively. The incidence rates in all age groups (at 5-year intervals) were higher in 2004 than in 1987/1988, indicating that people 50 years of age or older became more susceptible to hip fractures. Accordingly, the accretion of the hip fracture incidence rate was greater than that which could be explained purely by changes in population size and structure.
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Reliability, validity, and responsiveness of the Japanese version of the Patient-Rated Wrist Evaluation.
J Orthop Sci
PUBLISHED: 03-09-2010
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The Patient-Rated Wrist Evaluation is a regionspecific, self-administered questionnaire consisting of a pain scale (PRWE-P) and a functional scale (PRWE-F), with the latter consisting of specific function (PRWE-SF) and usual function (PRWE-UF). The PRWE was cross-culturally adapted from the original English version by the Impairment Evaluation Committee, Japanese Society for Surgery of the Hand (JSSH). The purpose of this study was to test the reliability, validity, and responsiveness of the Japanese version of PRWE (PRWE-J).
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In vivo gene transfer between interacting human osteosarcoma cell lines is associated with acquisition of enhanced metastatic potential.
J. Cell. Biochem.
PUBLISHED: 07-23-2009
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We report here in vivo gene transfer between cancer cells is associated with acquisition of high metastatic behavior. The 143B-GFP cell line with high metastatic potential and the MNNG/HOS-RFP cell line with low metastatic potential, both derived from the TE85 human osteosarcoma cell line, were either co-transplanted or transplanted alone in the tibia in nude mice. Upon mixed transplantation of the two differently labeled sublines, resulting metastatic colonies are single colored either red or green, thereby demonstrating their clonality and enabling facile color-coded quantification. When MNNG/HOS-RFP and 143B-GFP were co-transplanted in the tibia, the number of lung metastases of MNNG/HOS-RFP increased eight-fold compared to MNNG/HOS-RFP transplanted alone (P < 0.01). In contrast, no enhancement of MNNG/HOS-RFP metastases occurred when MNNG/HOS-RFP and 143B-GFP were transplanted separately in the right and left tibiae, respectively. This result suggests that the presence of 143B-GFP increased the metastatic potential of MNNG/HOS-RFP within the mixed tumor. We observed transfer of the Ki-ras gene from 143B-GFP to MNNG/HOS-RFP after they were co-implanted suggesting the Ki-ras played a role in increasing the metastatic potential of MNNG/HOS-RFP in the presence of 143B-GFP. These data suggest the possible role of in vivo gene transfer in enhancing the metastatic potential of cancer cells. The data also further demonstrated the power of color-coded imaging to visualize cancer-cell/cancer-cell interactions in vivo.
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A novel double-network hydrogel induces spontaneous articular cartilage regeneration in vivo in a large osteochondral defect.
Macromol Biosci
PUBLISHED: 06-13-2009
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We have developed a novel method to induce spontaneous hyaline cartilage regeneration in vivo for a large osteochondral defect by implanting a plug made from a double-network hydrogel composed of poly(2-acrylamido-2-methylpropanesulfonic acid) and poly(N,N-dimethylacrylamide) at the bottom of the defect, leaving the cavity vacant. In cells regenerated in the treated defect, type-2 collagen, Aggrican, and SOX9 mRNAs were highly expressed and the regenerated matrix was rich in proteoglycan and type-2 collagen at 4 weeks. This fact gave a significant modification to the commonly established concept that hyaline cartilage tissue cannot regenerate in vivo. This study prompted an innovative strategy in the field of joint surgery to repair an osteochondral defect using an advanced, high-function hydrogel.
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Vascularized fibular graft for bone defects after wide resection of musculoskeletal tumors.
J Orthop Sci
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In reconstruction by vascularized fibular graft (VFG) after wide resection of musculoskeletal tumors, there are problems such as the method of fixing the fibular graft, the period of achieving bone union, and the avoidance of postoperative fractures. We have performed VFG on 19 cases over a 30-year period. We have investigated these problems and now report the results.
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Will preoperative atrophy and Fatty degeneration of the shoulder muscles improve after rotator cuff repair in patients with massive rotator cuff tears?
Adv Orthop
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Recently, retear rate after repair for massive cuff tear have been improved through devised suture techniques. However, reported retear rate is relevant to preoperative atrophy and fatty degeneration. The purpose of this study was to investigate whether preoperative atrophy and fatty degeneration of rotator cuff muscles improve by successful repair. Twenty-four patients with massive rotator cuff tear were evaluated on the recovery of atrophy and fatty degeneration of supraspinatus and infraspinatus muscle after surgery. Atrophy was classified by the occupation ratio and fatty degeneration by modified Goutalliers classification. Both were assessed on magnetic resonance imaging (MRI) before and after the operation. When the cuff was well repaired, improvement of the atrophy and fatty degeneration were observed in a half and a one-fourth of the cases, respectively. In retear cases, however, atrophy and fatty degeneration became worse. Improvement of atrophy and fatty degeneration of the rotator cuff muscles may be expected in the cases with successful achievement of rotator cuff repair for large and massive tear.
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What is Visualize?

JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

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In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.