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1.
Vet Anaesth Analg ; 51(3): 266-270, 2024.
Article in English | MEDLINE | ID: mdl-38565449

ABSTRACT

OBJECTIVE: To develop and evaluate a low-cost three-dimensional (3D)-printed video laryngoscope (VLVET) for use with a commercial borescope. STUDY DESIGN: Instrument development and pilot study. ANIMALS: A total of six adult male Beagle dogs. METHODS: The VLVET consisted of a laryngoscope handle and a Miller-type blade, and a detachable camera holder that attached to various locations along the blade. The laryngoscope and camera holder were 3D-printed using black polylactic acid filament. Dogs were premedicated with intravenous (IV) medetomidine (15 µg kg-1) and anesthesia induced with IV alfaxalone (1.5 mg kg-1). The VLVET, combined with a borescope, was used for laryngeal visualization and intubation. Performance was evaluated by comparing direct and video-assisted views in sternal recumbency. The borescope camera was sequentially positioned at 2, 4, 6, 8 and 10 cm from the blade tip (distanceLARYNX-CAM), which was placed on the epiglottis during intubation or laryngoscopy. At the 10 cm distanceLARYNX-CAM, laryngeal visualization was sequentially scored at inter-incisor gaps of 10, 8, 6, 4 and 2 cm. Laryngeal visualization scores (0-3 range, with 0 = obstructed and 3 = unobstructed views) were statistically analyzed using the Friedman's test. RESULTS: Under direct visualization, the 2 cm distanceLARYNX-CAM had a significantly lower score compared with all other distanceLARYNX-CAM (all p = 0.014) because the view was obstructed by the camera holder and borescope camera. With both direct and camera-assisted views, visualization scores were higher at inter-incisor gaps ≥ 4 cm compared with 2 cm (all p < 0.05). CONCLUSIONS AND CLINICAL RELEVANCE: During laryngoscopy and intubation, the VLVET and borescope facilitated both direct and video laryngoscopy at distanceLARYNX-CAM in Beagle dogs when inter-incisor gaps were ≥ 4 cm.


Subject(s)
Intubation, Intratracheal , Laryngoscopes , Printing, Three-Dimensional , Animals , Dogs , Laryngoscopes/veterinary , Male , Intubation, Intratracheal/veterinary , Intubation, Intratracheal/instrumentation , Intubation, Intratracheal/methods , Video Recording , Laryngoscopy/veterinary , Laryngoscopy/methods , Laryngoscopy/instrumentation , Pilot Projects , Equipment Design
2.
Antioxidants (Basel) ; 13(1)2024 Jan 16.
Article in English | MEDLINE | ID: mdl-38247533

ABSTRACT

The process of cellular senescence, which is characterized by stable cell cycle arrest, is strongly associated with dysfunctional cellular metabolism and circadian rhythmicity, both of which are reported to result from and also be causal to cellular senescence. As a result, modifying any of them-senescence, metabolism, or the circadian clock-may affect all three simultaneously. Obesity accelerates aging by disrupting the homeostasis of reactive oxygen species (ROS) via an increased mitochondrial burden of fatty acid oxidation. As a result, if senescence, metabolism, and circadian rhythm are all linked, anti-obesity treatments may improve metabolic regulation while also alleviating senescence and circadian rhythm. Vutiglabridin is a small molecule in clinical trials that improves obesity by enhancing mitochondrial function. We found that chronic treatment of senescent primary human dermal fibroblasts (HDFs) with vutiglabridin alleviates all investigated markers of cellular senescence (SA-ß-gal, CDKN1A, CDKN2A) and dysfunctional cellular circadian rhythm (BMAL1) while remarkably preventing the alterations of mitochondrial function and structure that occur during the process of cellular senescence. Our results demonstrate the significant senescence-alleviating effects of vutiglabridin, specifically with the restoration of cellular circadian rhythmicity and metabolic regulation. These data support the potential development of vutiglabridin against aging-associated diseases and corroborate the intricate link between cellular senescence, metabolism, and the circadian clock.

3.
Front Vet Sci ; 10: 1287185, 2023.
Article in English | MEDLINE | ID: mdl-38026615

ABSTRACT

Primary renal neoplasia is rare in humans and dogs, with renal cell carcinoma (RCC) being the most common form of this cancer. As RCC is often diagnosed at an advanced stage, pulmonary metastasis is frequently observed. Tyrosine kinase inhibitors (TKIs) are the standard adjuvant treatments for metastatic RCC in humans. Similarly, in veterinary medicine, recent trials have employed TKIs for early-stage RCC patients who underwent complete surgical resection and showed no distant metastasis. However, the use of TKIs has not yet been reported commonly in cases of advanced RCC with metastasis. This case study presents the first clinical outcomes of TKI therapy in a dog with incompletely resected RCC and metastasis. A 5-year-old spayed female Chihuahua was referred to our hospital with a right renal mass and multiple pulmonary nodules suspected to be metastases. A portion of the renal mass was surgically removed, and histopathological examination revealed RCC with a high mitotic index. Adjuvant chemotherapy was administered, owing to incomplete resection with suspected pulmonary metastasis. An anticancer drug response prediction test was conducted using patient tissues. Since toceranib showed the most favorable responsiveness, it was selected as a therapeutic agent. Toceranib was orally administered at a dosage of 2.27 mg/kg every 48 h. Regular medical records for potential adverse effects were obtained, including systemic blood pressure, complete blood count, serum biochemical examination, and urinalysis. After 2 weeks of toceranib therapy, partial remission of pulmonary nodules continued for 2 months. The patient did not experience any adverse effects of the anticancer drug during the 4-month follow-up period. However, the patient died from an unidentified cause 6 months after the initial detection of the renal mass. This report describes the use of toceranib in dogs with RCC. In the present case, the patient showed an initial response to chemotherapy, and despite the presence of several poor prognostic factors, the dog survived beyond the expected 3-month lifespan to 6 months. Notably, no adverse events were observed during treatment.

4.
Exp Gerontol ; 181: 112269, 2023 10 01.
Article in English | MEDLINE | ID: mdl-37567452

ABSTRACT

BACKGROUND: Ageing alters the ECM, leading to mitochondrial dysfunction and oxidative stress, which triggers an inflammatory response that exacerbates with age. Age-related changes impact satellite cells, affecting muscle regeneration, and the balance of proteins. Furthermore, ageing causes a decline in NAD+ levels, and alterations in fat metabolism that impact our health. These various metabolic issues become intricately intertwined with ageing, leading to a variety of individual-level diseases and profoundly affecting individuals' healthspan. Therefore, we hypothesize that vutiglabridin capable of alleviating these metabolic abnormalities will be able to ameliorate many of the problems associated with ageing. METHOD: The efficacy of vutiglabridin, which alleviates metabolic issues by enhancing mitochondrial function, was assessed in aged mice treated with vutiglabridin and compared to untreated elderly mice. On young mice, vutiglabridin-treated aged mice, and non-treated aged mice, the Senescence-associated beta-galactosidase staining and q-PCR for ageing marker genes were carried out. Bulk RNA-seq was carried out on GA muscle, eWAT, and liver from each group of mice to compare differences in gene expression in various gene pathways. Blood from each group of mice was used to compare and analyze the ageing lipid profile. RESULTS: SA-ß-gal staining of eWAT, liver, kidney, and spleen of ageing mice showed that vutiglabridin had anti-ageing effects compared to the control group, and q-PCR of ageing marker genes including Cdkn1a and Cdkn2a in each tissue showed that vutiglabridin reduced the ageing process. In aged mice treated with vutiglabridin, GA muscle showed improved homeostasis compared to controls, eWAT showed restored insulin sensitivity and prevented FALC-induced inflammation, and liver showed reduced inflammation levels due to prevented TLO formation, improved mitochondrial complex I assembly, resulting in reduced ROS formation. Furthermore, blood lipid analysis revealed that ageing-related lipid profile was relieved in ageing mice treated with vutiglabridin versus the control group. CONCLUSION: Vutiglabridin slows metabolic ageing mechanisms such as decreased insulin sensitivity, increased inflammation, and altered NAD+ metabolism in adipose tissue in mice experiments, while also retaining muscle homeostasis, which is deteriorated with age. It also improves the lipid profile in the blood and restores mitochondrial function in the liver to reduce ROS generation.


Subject(s)
Insulin Resistance , Mice , Animals , Reactive Oxygen Species/metabolism , NAD , Aging/metabolism , Inflammation/genetics
5.
Vet Sci ; 10(2)2023 Feb 13.
Article in English | MEDLINE | ID: mdl-36851454

ABSTRACT

A 4-year-old mixed breed dog and a 19-year-old English cocker spaniel dog were evaluated for fecal incontinence. The second dog's fecal incontinence was associated with the anal mass. In both dogs, reconstruction of the external anal sphincter was required to gain fecal continence. Especially in the dog with an anal mass, the whole musculature involved in fecal continence was removed with the affected anorectum. Conventional surgical treatments for fecal incontinence have limitations in terms of muscle flap length and complexity of the surgical procedure. A modified surgical technique using the semitendinosus muscle was devised in the present study to overcome these limitations. The distal part of the semitendinosus muscle was bifurcated to make two muscle bundles, used to completely encircle the anorectum. These muscle bundles were sutured to the surrounding rectal muscle and the pelvic diaphragm to simulate the function of the external anal sphincter. Three months after surgery, both dogs showed significantly improved fecal continence without severe complications, such as infection, dehiscence, or lameness of the limb where the semitendinosus muscle was harvested. The outcomes of the two dogs supported the acceptability of the bifurcated muscle flap for anal sphincter augmentation. In addition, this report showed the possibility of more diverse applications of semitendinosus muscle in dogs.

6.
Biochem Biophys Res Commun ; 632: 62-68, 2022 12 03.
Article in English | MEDLINE | ID: mdl-36201882

ABSTRACT

Age-related macular degeneration (AMD) is a chronic and progressive disease characterized by degeneration of the retinal pigment epithelium (RPE) and retina that ultimately leads to loss of vision. The pathological mechanisms of AMD are not fully known. Cellular senescence, which is a state of cell cycle arrest induced by DNA-damage or aging, is hypothesized to critically affect the pathogenesis of AMD. In this study, we examined the relationship between cellular senescence and RPE/retinal degeneration in mouse models of natural aging and accelerated aging. We performed a bulk RNA sequencing of the RPE cells from adult (8 months old) and naturally-aged old (24 months old) mice and found that common signatures of senescence and AMD pathology - inflammation, apoptosis, and blood vessel formation - are upregulated in the RPE of old mice. Next, we investigated markers of senescence and the degree of RPE/retinal degeneration in Zmpste24-deficient (Zmpste24-/-) mice, which is a model for progeria and accelerated aging. We found that Zmpste24-/- mice display markedly greater level of senescence-related markers in RPE and significant RPE/retinal degeneration compared to wild-type mice, in a manner consistent with natural aging. Overall, these results provide support for the association between cellular senescence of RPE and the pathogenesis of AMD, and suggest the use of Zmpste24-/- mice as a novel senescent RPE model of AMD.


Subject(s)
Macular Degeneration , Retinal Degeneration , Retinal Pigment Epithelium , Animals , Mice , Aging/pathology , DNA/metabolism , Macular Degeneration/genetics , Macular Degeneration/pathology , Membrane Proteins/genetics , Membrane Proteins/metabolism , Metalloendopeptidases/genetics , Metalloendopeptidases/metabolism , Phenotype , Retinal Degeneration/genetics , Retinal Degeneration/pathology , Retinal Pigment Epithelium/metabolism
7.
Adv Healthc Mater ; 5(2): 205-12, 2016 Jan 21.
Article in English | MEDLINE | ID: mdl-26638984

ABSTRACT

Here, a growth-factor-integrated natural extracellular matrix of type I collagen is presented that induces angiogenesis. The developed matrix adapts type I collagen nanofibers integrated with synthetic colloidal particles of recombinant bacteriophages that display vascular endothelial growth factor (VEGF). The integration is achieved during or after gelation of the type I collagen and the matrix enables spatial delivery of VEGF into a desired region. Endothelial cells that contact the VEGF are found to invade into the matrix to form tube-like structures both in vitro and in vivo, proving the angiogenic potential of the matrix.


Subject(s)
Bacteriophages/metabolism , Collagen Type I/pharmacology , Extracellular Matrix/metabolism , Neovascularization, Physiologic/drug effects , Recombination, Genetic/genetics , Vascular Endothelial Growth Factors/metabolism , Animals , Cell Surface Display Techniques , Enzyme-Linked Immunosorbent Assay , Extracellular Matrix/drug effects , Fluorescent Antibody Technique , Humans , Mice, Inbred BALB C , Microscopy, Fluorescence , Nanofibers/chemistry , Protein Engineering , Rats
8.
Chem Commun (Camb) ; 50(82): 12392-5, 2014 Oct 21.
Article in English | MEDLINE | ID: mdl-25188309

ABSTRACT

Titanium-tungsten oxide composites with greatly enhanced photocatalytic activity were synthesized by lysozyme-mediated biomineralization. It was shown for the first time that simple control of the onset of biomineralization could enable fine tuning of the composition and crystallinity of the composites to determine their photocatalytic performance.


Subject(s)
Muramidase/chemistry , Nanoparticles/chemistry , Oxides/chemistry , Titanium/chemistry , Tungsten/chemistry , Catalysis , Cellulose/analogs & derivatives , Cellulose/chemistry , Nanoparticles/radiation effects , Oxides/radiation effects , Photochemical Processes , Rhodamines/chemistry , Sunlight , Titanium/radiation effects , Tungsten/radiation effects
9.
Biomed Mater ; 9(1): 015009, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24448635

ABSTRACT

This paper reports a novel approach for the formation of anti-inflammatory surface coating on a neural electrode. The surface coating is realized using a recombinant f88 filamentous bacteriophage, which displays a short platinum binding motif and a tumor necrosis factor alpha antagonist (TNF-α antagonist) on p3 and p8 proteins, respectively. The recombinant bacteriophages are immobilized on the platinum surface by a simple dip coating process. The selective and stable immobilization of bacteriophages on a platinum electrode is confirmed by quartz crystal microbalance with dissipation monitoring, atomic force microscope and fluorescence microscope. From the in vitro cell viability test, the inflammatory cytokine (TNF-α) induced cell death was prevented by presenting recombinant bacteriophage coating, albeit with no significant cytotoxic effect. It is also observed that the bacteriophage coating does not have critical effects on the electrochemical properties such as impedance and charge storage capacities. Thus, this approach demonstrates a promising anti-apoptotic as well as anti-inflammatory surface coating for neural implant applications.


Subject(s)
Bacteriophages/genetics , Drug Delivery Systems , Genetic Engineering/methods , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Amino Acid Motifs , Anti-Inflammatory Agents/chemistry , Apoptosis , Base Sequence , Cell Line, Tumor , Cell Survival , DNA/chemistry , Elasticity , Electrodes , Humans , Microscopy, Fluorescence , Molecular Sequence Data , Peptides/chemistry , Platinum/chemistry , Surface Properties , Tumor Necrosis Factor-alpha/chemistry , Viscosity
10.
J Nanosci Nanotechnol ; 13(9): 6203-7, 2013 Sep.
Article in English | MEDLINE | ID: mdl-24205629

ABSTRACT

Biological materials with surface-active proteins can be genetically modified to bind target materials. In particular, filamentous-shaped M13 bacteriophages (M13 phage) are attractive scaffolds for functional nanostructures due to their highly ordered protein-coat surface. This paper demonstrates a simple method for fabricating silica nanocables along a modified M13 phage. The M13 phage was genetically engineered to display the amino acid serine on the surface to provide hydroxyl groups for a sol-gel reaction. This M13 phage mutant offers homogeneous molecular templates for forming silica coated coaxial nanocables. Silica shell formation was confirmed by transmission electron microscopy (TEM) and electron dispersive X-ray (EDX) analysis. The core-shell structures were clearly distinguishable in the TEM analysis, and the synthesized shells were observed by EDX analysis. In addition, we investigated the adsorption properties of M13 phages on the pretreated substrate as a function of concentration. The effect of the relative concentration of M13 phages on the substrate was observed by using atomic force microscopy (AFM). We also fabricated top electrodes on the extremely dense network for measuring electrical properties of the M13 phage. The experimental DC measurement indicated that the wild-type phage has very low electrical conductance, similar to insulating material.


Subject(s)
Bacteriophage M13/chemistry , Nanostructures , Silicon Dioxide/chemistry , Microscopy, Electron, Transmission , Spectrometry, X-Ray Emission
11.
Sensors (Basel) ; 13(1): 455-62, 2012 Dec 27.
Article in English | MEDLINE | ID: mdl-23271605

ABSTRACT

Bacteriorhodopsin protein (bR)-based systems are one of the simplest known biological energy converters. The robust chemical, thermal and electrochemical properties of bR have made it an attractive material for photoelectric devices. This study demonstrates the photoelectric response of a dry bR layer deposited on a nitrocellulose membrane with indium tin oxide (ITO) electrodes. Light-induced electrical current as well as potential and impedance changes of dried bR film were recorded as the function of illumination. We have also tested bR in solution and found that the electrical properties are strongly dependent on light intensity changing locally proton concentration and thus pH of the solution. Experimental data support the assumption that bR protein on a positively charged nitrocellulose membrane (PNM) can be used as highly sensitive photo- and pH detector. Here the bR layer facilitates proton translocation and acts as an ultrafast optoelectric signal transducer. It is therefore useful in applications related to bioelectronics, biosensors, bio-optics devices and current carrying junction devices.


Subject(s)
Bacteriorhodopsins/metabolism , Collodion/metabolism , Electrochemistry/methods , Halobacterium salinarum/metabolism , Photochemistry/methods , Purple Membrane/metabolism , Electric Impedance , Hydrogen-Ion Concentration , Light , Purple Membrane/radiation effects
12.
Analyst ; 137(3): 575-9, 2012 Feb 07.
Article in English | MEDLINE | ID: mdl-22189821

ABSTRACT

In this work we present a label free quantitative detection method for DNA samples amplified by polymerase chain reaction (PCR) in aqueous medium using terahertz-time domain spectroscopy (THz-TDS) in the frequency range from 0.3 to 1.2 THz. The DNA samples of 133 and 697 base pairs were prepared using PCR. We measured the absorption coefficients of DNA solutions in the concentration range of 0-0.3 ng µl(-1). For both DNA types, the absorption coefficients decreased with increasing DNA concentrations. The average change in absorption coefficients compared to buffer within the frequency range of 0.8-1.0 THz showed a linear behavior. Our results demonstrate that THz-TDS can detect PCR amplified DNA in aqueous solution with a minimum concentration of 0.1 ng µl(-1) and a minimum sample volume of 10 µl.


Subject(s)
DNA/analysis , Polymerase Chain Reaction/methods , Spectrum Analysis/methods , Water/chemistry , Base Sequence , DNA Primers , Solutions
13.
Cancer Res ; 69(11): 4784-90, 2009 Jun 01.
Article in English | MEDLINE | ID: mdl-19487290

ABSTRACT

LMO2 is a transcription regulator involved in human T-cell leukemia, including some occurring in X-SCID gene therapy trials, and in B-cell lymphomas and prostate cancer. LMO2 functions in transcription complexes via protein-protein interactions involving two LIM domains and causes a preleukemic T-cell development blockade followed by clonal tumors. Therefore, LMO2 is necessary but not sufficient for overt neoplasias, which must undergo additional mutations before frank malignancy. An open question is the importance of LMO2 in tumor development as opposed to sustaining cancer. We have addressed this using a peptide aptamer that binds to the second LIM domain of the LMO2 protein and disrupts its function. This specificity is mediated by a conserved Cys-Cys motif, which is similar to the zinc-binding LIM domains. The peptide inhibits Lmo2 function in a mouse T-cell tumor transplantation assay by preventing Lmo2-dependent T-cell neoplasia. Lmo2 is, therefore, required for sustained T-cell tumor growth, in addition to its preleukemic effect. Interference with LMO2 complexes is a strategy for controlling LMO2-mediated cancers, and the finger structure of LMO2 is an explicit focus for drug development.


Subject(s)
Aptamers, Peptide/pharmacology , DNA-Binding Proteins/antagonists & inhibitors , DNA-Binding Proteins/physiology , Drug Delivery Systems/methods , Metalloproteins/antagonists & inhibitors , Metalloproteins/physiology , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/genetics , Adaptor Proteins, Signal Transducing , Amino Acid Sequence , Animals , Aptamers, Peptide/metabolism , Aptamers, Peptide/therapeutic use , CHO Cells , Cricetinae , Cricetulus , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Humans , LIM Domain Proteins , Metalloproteins/genetics , Metalloproteins/metabolism , Mice , Mice, Transgenic , Models, Molecular , Molecular Sequence Data , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/drug therapy , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/pathology , Protein Binding/genetics , Protein Interaction Domains and Motifs/genetics , Tumor Cells, Cultured , Zinc Fingers/genetics , Zinc Fingers/physiology
14.
Biosens Bioelectron ; 24(5): 1324-9, 2009 Jan 01.
Article in English | MEDLINE | ID: mdl-19036573

ABSTRACT

Generally, an immunoaffinity SPR biosensor detects a target analyte in a sample through highly selective adsorption by using the antigen-antibody interaction. For improving the sensitivity, various kinds of particles have been added to the already bound analytes on the SPR biosensor (sandwich assay). In this work, signal amplification was demonstrated by the expression of the IgG-binding Z-domain of protein A on the outer membrane of Escherichia coli via "Autodisplay". The amount of Z-domain of protein A expressed on the outer membrane was calculated to be 280,000 molecules per cell. In addition, the IgG-binding ability of the expressed protein was characterized using FACS analysis. The signal amplification of the SPR biosensor was performed in the sandwich assay format using a model of horseradish peroxidase (HRP); the limit of detection was determined to be significantly improved from 1 microg/ml to 1 ng/ml. Finally, myoglobin analysis was demonstrated for the medical diagnosis of cardiac diseases. The detection limit was estimated to be improved from 10 ng/ml to <1 ng/ml. These results show that Z-domain-displaying E. coli can be successfully used for the signal amplification of immunoaffinity biosensors, thereby improving the sensitivity and the limit of detection.


Subject(s)
Biological Assay/instrumentation , Biosensing Techniques/instrumentation , Escherichia coli/metabolism , Immunoassay/instrumentation , Immunoglobulin G/analysis , Staphylococcal Protein A/metabolism , Surface Plasmon Resonance/instrumentation , Biosensing Techniques/methods , Equipment Design , Equipment Failure Analysis , Escherichia coli/drug effects , Escherichia coli/genetics , Immunoglobulin G/metabolism , Protein Engineering/methods , Protein Structure, Tertiary , Reproducibility of Results , Sensitivity and Specificity , Staphylococcal Protein A/genetics
15.
Mol Ther ; 13(1): 15-25, 2006 Jan.
Article in English | MEDLINE | ID: mdl-16260184

ABSTRACT

Chromosomal translocations are primary events in the development of leukemias, representing at least one genetic feature of the putative cancer stem cell. Studies of genes influenced by chromosomal translocations have yielded a vast amount of information about how cancer is initiated and maintained. In particular, acute leukemias have demonstrated that chromosomal translocations often involve transcription regulators that function by interacting with proteins and by controlling cell fate in the aberrant setting of the developing cancer cell. As a quintessential chromosomal translocation gene product, LMO2 has many properties that typify this class of molecule. In addition to its involvement in chromosomal translocations, the LMO2 gene was inadvertently activated in an X-SCID gene therapy trial by retroviral insertion. New molecular therapies targeted directly at the LMO2 protein could have major impact as adjuncts to existing therapies or as therapeutics in their own right. In this review, we outline the current knowledge about LMO2 and some possible routes to develop reagents that might be possible macromolecular drugs in the future.


Subject(s)
DNA-Binding Proteins/physiology , Leukemia, T-Cell/metabolism , Metalloproteins/physiology , Retroviridae/genetics , Transcription, Genetic , Translocation, Genetic/physiology , Adaptor Proteins, Signal Transducing , Animals , DNA-Binding Proteins/genetics , Endothelium, Vascular/physiology , Genetic Therapy , Hematopoiesis , Humans , LIM Domain Proteins , Leukemia, T-Cell/genetics , Metalloproteins/genetics , Mice , Mice, Transgenic , Multiprotein Complexes/physiology , Neovascularization, Pathologic/genetics , Neovascularization, Pathologic/metabolism , Proto-Oncogene Proteins , Translocation, Genetic/genetics
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