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Exosomes are potent players in the development of metastases and they play an important role in cancer angiogenesis and exacerbation. However, it is unclear how proteins on exosomes affect development of blood vessel networks. In this study, we focused on relationships between membrane proteins on exosomes and angiogenesis using human umbilical vein endothelial cells (HUVEC). Lung tumor cell-derived exosomes induced tube formation and growth of endothelial cells in vitro in a dose-dependent manner involving MAPK activation, but this was not seen in normal lung epithelial cells. Ephrin type-A receptor 2 (EphA2) was identified by proteomic analysis and an inhibition assays showed it is a major MAPK activator on exosomes. Thus EphA2 on exosomes participates in angiogenesis as a ligand of the ephrin signaling pathway. These results support the development of novel therapeutic strategies such as blockade of remote cancer communications through exosomes.
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Efrina-A2/metabolismo , Exosomas/metabolismo , Neoplasias Pulmonares/irrigación sanguínea , Sistema de Señalización de MAP Quinasas , Inductores de la Angiogénesis , Línea Celular Tumoral , Células Epiteliales/metabolismo , Células Endoteliales de la Vena Umbilical Humana , Humanos , Neoplasias Pulmonares/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Neovascularización Patológica/metabolismo , Neovascularización Patológica/patología , Cultivo Primario de Células , Receptor EphA2 , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismoRESUMEN
The isospin character of p-n pairs at large relative momentum has been observed for the first time in the ^{16}O ground state. A strong population of the J,T=1,0 state and a very weak population of the J,T=0,1 state were observed in the neutron pickup domain of ^{16}O(p,pd) at 392 MeV. This strong isospin dependence at large momentum transfer is not reproduced by the distorted-wave impulse approximation calculations with known spectroscopic amplitudes. The results indicate the presence of high-momentum protons and neutrons induced by the tensor interactions in the ground state of ^{16}O.
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AIMS: The aim of this study was to evaluate the correlation between Salter's criteria and Kalamchi's classification of avascular necrosis in patients treated for developmental dysphasia of the hip (DDH). PATIENTS AND METHODS: The study involved a retrospective analysis of 123 patients (123 hips) with DDH treated by operative and non-operative reduction before the age of two years, with a minimum follow-up of ten years. Salter's criteria (S1 to S4) were determined from radiographs obtained at one to two years post-reduction, whilst the Kalamchi grade was determined from radiographs obtained at ten or more years of age. Early post-reduction radiographs were also used to evaluate the centre-head distance discrepancy (CHDD) and the occurrence of a dome-shaped deformity of the proximal femoral metaphysis (D-shaped metaphysis). The prognosis was described as good (Kalamchi grade K0 or KI), fair (Kalamchi grade KII) or poor (Kalamchi grade KIII or KIV) for analysis and correlation with the early Salter criteria, CHDD and D-shaped metaphysis. RESULTS: S1 and S2 criteria were predictive of a poor prognosis. The outcome following S3, S4 and S3 + S4 varied; 18 (40%) had a good prognosis, 17 (38%) a fair prognosis and ten (22%) a poor prognosis. A CHDD ≥ 10% and a D-shaped metaphysis were also predictive of a poor prognosis. CONCLUSION: The Salter criteria were predictive of the Kalamchi grade of avascular necrosis in patients with DDH aged ten or more years after reduction of the hip. Cite this article: Bone Joint J 2017;99-B:1115-20.
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Necrosis de la Cabeza Femoral/diagnóstico , Luxación Congénita de la Cadera/cirugía , Osteotomía/efectos adversos , Complicaciones Posoperatorias , Niño , Preescolar , Femenino , Necrosis de la Cabeza Femoral/epidemiología , Necrosis de la Cabeza Femoral/etiología , Estudios de Seguimiento , Humanos , Incidencia , Japón/epidemiología , Masculino , Pronóstico , Radiografía , Estudios Retrospectivos , Factores de TiempoRESUMEN
BACKGROUND AND OBJECTIVES: Human leukocyte antigen (HLA) antibodies, which are involved in the development of transfusion-related side effects such as transfusion-related lung injury, are sometimes found in males without a history of alloimmunization (eg, transplantation and transfusion). Whether HLA antibodies in male donors can interact with their target HLA specificities expressed on cells have not been completely investigated. MATERIALS AND METHODS: The HLA antibodies detected in 7 male donors were characterized. Flow cytometry and immunocomplex capture fluorescence analysis were performed to evaluate the ability of these antibodies to bind with target HLA specificities expressed on cells. The association of these antibodies with complement was examined using anti-C1q antibody. Sustainability of HLA antibodies over time was compared in 26 male vs 57 female donors. RESULTS: The antibodies from all 7 donors recognized intact HLA molecules coated onto microbeads. The antibodies in 2 of 7 donors also recognized their target HLA specificities expressed on cells. Furthermore, the antibodies in one of these 2 donors showed HLA specificities that involved complement binding. Twenty-one of 26 initially positive male donors had turned negative for HLA antibody at least 1 year after their initial positive screening, whereas HLA antibody positivity was maintained for a long time in most female donors. CONCLUSION: Males without apparent alloimmunization could have HLA antibodies that recognize their target HLA specificities on cells and that could potentially modify molecular events in affected cells.
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Complejo Antígeno-Anticuerpo/sangre , Donantes de Sangre , Proteínas del Sistema Complemento/metabolismo , Antígenos HLA/sangre , Isoanticuerpos/sangre , Adulto , Especificidad de Anticuerpos , Femenino , Citometría de Flujo , Humanos , Masculino , Unión Proteica , Factores Sexuales , Lesión Pulmonar Aguda Postransfusional/prevención & controlRESUMEN
Tumor necrosis factor (TNF)/TNF receptors (TNFR1/TNFR2) are considered to be potential drug targets to treat refractory diseases, including autoimmune diseases and malignant tumors. However, their specific functions, especially in the case of TNFR2, are poorly understood. In this study, we constructed a mouse TNFR2 (mTNFR2)-mediated biological assay system that shows no effects of mouse TNFR1 (mTNFR1) in order to screen mTNFR2-selective stimulating agents. Mouse TNFR1(-/-)R2(-/-) preadipocytes were transfected with the gene encoding the mTNFR2/mouse Fas (mFas) chimeric receptor in which the extracellular and transmembrane domains of mTNFR2 were fused to the intracellular domain of mFas. Our results demonstrated that this cell line exhibits highly sensitive mTNFR2-mediated cytotoxic effects. We propose that this mTNFR2-mediated biological assay system would be a useful tool to screen for mTNFR2-selective stimulating agents.
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Adipocitos/citología , Receptores Tipo II del Factor de Necrosis Tumoral/genética , Receptores Tipo I de Factores de Necrosis Tumoral/genética , Receptor fas/genética , Animales , Bioensayo/métodos , Línea Celular , Ratones , Ratones Noqueados , Receptores Tipo I de Factores de Necrosis Tumoral/efectos de los fármacos , Receptores Tipo II del Factor de Necrosis Tumoral/efectos de los fármacos , TransfecciónRESUMEN
BACKGROUND: The relationship among natural allergen exposure, induction of blocking antibody and the occurrence of atopic allergy-particularly in the presence of IgE production-is debatable. OBJECTIVE: To clarify the relationship between the dose of cutaneous exposure to dust mite allergen and susceptibility to the IgE-mediated allergic response in relation to IgG production. METHODS: NC/Nga mice were epicutaneously exposed to various doses of Dermatophagoides pteronyssinus allergen to induce atopic dermatitis-like skin lesions. We then evaluated the skin lesions, induction of mite-specific immune responses, and susceptibility to anaphylaxis. RESULTS: Dose-dependent exacerbation of atopic dermatitis-like skin lesions and increases in mite-specific IgG and IgE production were observed. However, mice exposed to relatively low doses of mite allergen showed hypersusceptibility to mite allergen-specific anaphylaxis. We also showed that adoptive transfer of total IgG from Dp-sensitized mice rescued mice from the hypersusceptibility seen in those exposed to low doses of mite allergen. CONCLUSIONS AND CLINICAL RELEVANCE: High-dose cutaneous exposure to dust mites induced effective blocking IgG production, even if accompanied by IgE production. Our data might support the concept that an increase in IgG titre, not a decrease in IgE titre, is a marker of clinical improvement in allergen-specific immunotherapy.
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Alérgenos/administración & dosificación , Alérgenos/inmunología , Anafilaxia/prevención & control , Anticuerpos Bloqueadores/inmunología , Antígenos Dermatofagoides/administración & dosificación , Antígenos Dermatofagoides/inmunología , Inmunoglobulina G/inmunología , Anafilaxia/inmunología , Anafilaxia/metabolismo , Animales , Especificidad de Anticuerpos/inmunología , Citocinas/metabolismo , Modelos Animales de Enfermedad , Femenino , Inmunización , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Inmunoglobulina G/sangre , RatonesRESUMEN
The fullerene C60 is used in consumer products such as cosmetics owing to its antioxidative effects and is being developed for nanomedical applications. However, knowledge regarding the safety of fullerene C60, especially after oral administration, is sparse. Here, we examined the safety of fullerene C60 in mice after 7 d of exposure to orally administered polyvinylpyrrolidone (PVP)-wrapped fullerene C60 (PVP-fullerene C60). Mice treated with PVP-fullerene C60 showed few changes in the plasma levels of various markers of kidney and liver injury and experienced no significant hematologic effects. Furthermore, the histology of the colon of PVP-fullerene C60-treated mice was indistinguishable from that of control mice. These results suggest that PVP-fullerene C60 lacks toxicity after high-dose oral administration and indicate that PVP-fullerene C60 can be considered safe for oral medication. These data provide basic information that likely will facilitate the production of safe and effective forms of fullerene C60.
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Fulerenos/farmacología , Lesión Renal Aguda/inducido químicamente , Lesión Renal Aguda/patología , Administración Oral , Animales , Recuento de Células Sanguíneas , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Colitis/inducido químicamente , Colitis/patología , Femenino , Fulerenos/administración & dosificación , Luz , Ratones , Ratones Endogámicos C57BL , Povidona , Dispersión de Radiación , Fijación del TejidoRESUMEN
Detection of drug-target proteins and biomarkers that are expressed in cancer tissue has significant potential for both diagnosis and treatment of cancer. However, current immuno-histochemical and cytogenetic analyses of biopsy specimens for pre-operational diagnosis are highly invasive and often difficult to apply to lung cancer patients. The purpose of this study was to evaluate the possible utility of determining epidermal growth factor receptor (EGFR) expression on exosomal membranes using a targeted ELISA with an anti-CD81 antibody as a capture antibody for lung cancer diagnosis. While soluble EGFR (sEGFR) levels in plasma were not remarkably different between lung cancer patients and normal controls, significantly higher exosomal EGFR expression levels were observed in 5/9 cancer cases compared to normal controls. These results suggest that measurement of exosomal protein levels could be useful for in vitro diagnosis, and that exosomal EGFR is a possible biomarker for characterization of lung cancer.
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Biomarcadores/análisis , Receptores ErbB/metabolismo , Exosomas/metabolismo , Neoplasias Pulmonares/diagnóstico , Adulto , Anciano , Animales , Western Blotting , Línea Celular Tumoral , Medios de Cultivo Condicionados , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunohistoquímica , Masculino , Membranas/metabolismo , Ratones , Ratones Endogámicos BALB C , Microscopía Electrónica , Persona de Mediana Edad , Plásmidos , Tetraspanina 28/metabolismoRESUMEN
Since metastasis is one of the most important prognostic factors in colorectal cancer, development of new methods to diagnose and prevent metastasis is highly desirable. However, the molecular mechanisms leading to the metastatic phenotype have not been well elucidated. In this study, a proteomics-based search was carried out for metastasis-related proteins in colorectal cancer by analyzing the differential expression of proteins in primary versus metastasis focus-derived colorectal tumor cells. Protein expression profiles were determined using a tissue microarray (TMA), and the results identified Rho GDP-dissociation inhibitor alpha (Rho GDI) as a metastasis-related protein in colon and prostate cancer patients. Consequently, Rho GDI may be useful as a diagnostic biomarker and/or a therapeutic to prevent colon and prostate cancer metastasis.
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Neoplasias del Colon/secundario , Inhibidores de Disociación de Guanina Nucleótido/fisiología , Neoplasias de la Próstata/secundario , Anciano , Western Blotting , Línea Celular Tumoral , Cromatografía Líquida de Alta Presión , Electroforesis en Gel Bidimensional , Colorantes Fluorescentes , Geles , Gliceraldehído-3-Fosfato Deshidrogenasas/metabolismo , Humanos , Hidrólisis , Inmunohistoquímica , Masculino , Espectrometría de Masas , Análisis por Micromatrices , Persona de Mediana Edad , Tripsina/química , Inhibidores de la Disociación del Nucleótido Guanina rho-EspecíficoRESUMEN
Fetal fibroblast cell culture from cotyledons of bovine placenta and animal experiments close to term were used to elucidate afterbirth release and factors missing in the signal transduction mechanism for retained fetal membranes (RFM) after delivery. In cell culture the addition of arachidonic acid (Ara) to the medium caused rapid release to free floating cell in the culture dish, accompanied by matrix metalloproteinase (MMP) activation, being consistent with previous in vivo observations, where a relation between MMP and fetal membrane release had been shown. Ara-induced cell floating was not inhibited by the addition of cyclooxygenase (COX) inhibitor, and not induced by the addition of PGF2α or PGE2 to replace Ara, while 12-lipoxygenase (12-LOX) metabolite of Ara, 12-oxo-eicosatetraenoic acid (12-oxoETE), strongly induced cell floating. In the animal experiments, 12-oxoETE injection to delivery-induced cows (n = 6) using prostaglandin (PG) and dexamethazone resulted in rapid release of fetal membranes. In cows with natural calf delivery, a 12-oxoETE peak (11.7-16.8 ng/ml) was observed in maternal blood plasma prior to release of fetal membranes. This investigation thus gives new indications for that the mediator for fetal membrane release is 12-oxoETE and not PG.
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Ácidos Araquidónicos/farmacología , Membranas Extraembrionarias/efectos de los fármacos , Retención de la Placenta/veterinaria , Animales , Araquidonato 12-Lipooxigenasa/metabolismo , Bovinos , Células Cultivadas , Dexametasona/farmacología , Membranas Extraembrionarias/metabolismo , Femenino , Retención de la Placenta/metabolismo , Embarazo , Prostaglandinas/farmacologíaRESUMEN
The immune-modulating effect following intradermal injection of various-sized amorphous silica particles was analyzed in terms of induction of ovalbumin-specific CD8+ T cells in vivo. IFN-gamma ELISPOT assays revealed that only nanosilica particles with a diameter of less than 100 nm significantly enhanced CD8+ T cell responses against ovalbumin. These results indicate that the size of nanomaterials is a critical determinant in terms of their safe use.
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Factores Inmunológicos , Nanopartículas , Dióxido de Silicio/farmacología , Animales , Linfocitos T CD8-positivos/efectos de los fármacos , Linfocitos T CD8-positivos/inmunología , Femenino , Interferón gamma , Ratones , Ratones Endogámicos C57BL , Ovalbúmina/inmunología , Tamaño de la Partícula , Dióxido de Silicio/química , Bazo/citología , Bazo/inmunologíaRESUMEN
Adult T-cell leukemia (ATL) is a severe chemotherapy-resistant malignancy associated with prolonged infection by the human T cell-lymphotropic virus 1 (HTLV-1) retrovirus. Epidemiology studies strongly indicate that an increase in HTLV-1 virus load is an important factor during the onset of ATL. Therefore, inhibition of the growth/transmission of HTLV-1 infected cells is a promising strategy in preventing the disease. In our previous study, we revealed that arsenic trioxide (As2O3), a drug used to treat acute promyelocytic leukemia (APL), exerts an inhibitory effect on syncytium formation between HTLV-1 infected cells and HeLa cells via suppression of HTLV-1 envelope protein gp46 expression at low concentrations. In this study, we analyze the mechanism of action of As2O3 using a proteomics approach. Our results suggest that down-regulation of gp46 might be related to As2O3-induced oxidation of the 71-kDa heat shock cognate protein (HSC70) and the 78-kDa glucose-regulated protein (BiP/GRP78). We postulate that AS2O3 exerts an inhibitory effect on HTLV-1 virus transmission via down-regulation of gp46-production, which might be caused by oxidative modification of various proteins such as chaperones.
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Arsenicales/farmacología , Productos del Gen env/biosíntesis , Infecciones por HTLV-I/metabolismo , Óxidos/farmacología , Proteínas Oncogénicas de Retroviridae/biosíntesis , Trióxido de Arsénico , Fusión Celular , Regulación hacia Abajo/efectos de los fármacos , Electroforesis en Gel Bidimensional , Chaperón BiP del Retículo Endoplásmico , Geles , Productos del Gen env/antagonistas & inhibidores , Células HeLa , Humanos , Hidrólisis , Inmunoprecipitación , Oxidación-Reducción , Proteómica , Proteínas Oncogénicas de Retroviridae/antagonistas & inhibidores , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Tripsina/químicaRESUMEN
The purpose of this study was to establish a long-term tooth cryopreservation method that can be used for tooth autotransplantation. Human periodontal ligament (PDL) cells were frozen in 10% dimethyl sulfoxide (Me(2)SO) using a programmed freezer with a magnetic field. Cells were cryopreserved for 7 days at -150 degrees C. Immediately after thawing, the number of surviving cells was counted and the cells were cultured; cultured cells were examined after 48 h. Results indicated that a 0.01 mT of a magnetic field, a 15-min hold-time, and a plunging temperature of -30 degrees C led to the greatest survival rate of PDL cells. Based on these findings, whole teeth were cryopreserved under the same conditions for 1 year. The organ culture revealed that the PDL cells of cryopreserved tooth with a magnetic field could proliferate as much as a fresh tooth, although the cells did not appear in the cryopreserved tooth without a magnetic field. Histological examination and the transmission electron microscopic image of cryopreserved tooth with a magnetic field did not show any destruction of cryopreserved cells. In contrast, severe cell damage was seen in cells frozen without a magnetic field. These results indicated that a magnetic field programmed freezer is available for tooth cryopreservation.
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Criopreservación/métodos , Crioprotectores/farmacología , Ligamento Periodontal/citología , Bancos de Tejidos , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Dimetilsulfóxido/farmacología , Campos Electromagnéticos , Humanos , Magnetismo , Microscopía Electrónica de Transmisión , Técnicas de Cultivo de Órganos , Ligamento Periodontal/efectos de los fármacos , Ligamento Periodontal/efectos de la radiaciónRESUMEN
The development of a safe and effective mucosal vaccine adjuvant is a crucial step for the development of vaccines against human immunodeficiency virus type-1 (HIV). We have previously reported that a mutant tumor necrosis factor-alpha (TNF-alpha), mTNF-K90R, possessed strong mucosal vaccine adjuvant activities in mice. Here, we evaluated the potential of mTNF-K90R as a mucosal vaccine adjuvant for the induction of systemic and mucosal immune responses against HIV. Nasal immunization of BALB/c mice with 5 microg of an HIV gp120 env protein immunogen together with mTNF-K90R induced higher serum anti-HIV gp120 protein immunoglobulin G (IgG) responses than gp120 alone. Furthermore, mTNF-K90R induced anti-gp120 IgA responses in nasal as well as vaginal washes from immunized mice, although these were not administration sites. Again, responses with mTNF-K90R were higher than with gp120 alone. These results indicate that mTNF-K90R may be applicable as amucosal adjuvant for HIV vaccination to induce both systemic and mucosal immune responses.
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Vacunas contra el SIDA/genética , Vacunas contra el SIDA/inmunología , Adyuvantes Inmunológicos , Inmunidad Mucosa/inmunología , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunología , Animales , Ensayo de Inmunoadsorción Enzimática , Femenino , Proteína gp120 de Envoltorio del VIH/inmunología , Inmunización , Inmunoglobulina A/biosíntesis , Inmunoglobulina G/biosíntesis , Ratones , Ratones Endogámicos BALB C , Membrana Mucosa/inmunología , Ovalbúmina/inmunologíaRESUMEN
Amorphous silica nanoparticles (nSPs), are widely used in medicines, cosmetics and food. However, due to their reduced particle size they are suspected to pose new risks induced by changes in biological reactivity and kinetics, which differ from those of bulk materials. In a previous study, we showed that silica particles with a diameter of 70 nm penetrated the stratum corneum (SC) of mouse skin and were taken up by living cells such as keratinocytes and Langerhans cells. To clarify the relationship between particle size, distribution and cellular response, we have evaluated size-dependent intracellular localization and cytotoxicity of silica particles, using the mouse epidermal Langerhans cell line XS52. On treatment with silica particles of diameters 70, 300, and 1000 nm, cellular uptake and cytotoxicity increased with reduction in particle size. These results suggest that smaller sized silica particles induced greater cytotoxicity against Langerhans cells, which was correlated with the quantity of particle uptake into the cells.
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Células de Langerhans/efectos de los fármacos , Nanopartículas/toxicidad , Dióxido de Silicio/toxicidad , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Queratinocitos/efectos de los fármacos , L-Lactato Deshidrogenasa/metabolismo , Células de Langerhans/enzimología , Células de Langerhans/ultraestructura , Ratones , Microscopía Electrónica de Transmisión , Tamaño de la Partícula , Timidina/metabolismoRESUMEN
Tumor necrosis factor-alpha (TNF), which binds two types of TNF receptors (TNFR1 and TNFR2), regulates the onset and exacerbation of autoimmune diseases such as rheumatoid arthritis and Crohn's disease. In particular, TNFR1-mediated signals are predominantly related to the induction of inflammatory responses. We have previously generated a TNFR1-selective antagonistic TNF-mutant (mutTNF) and shown that mutTNF efficiently inhibits TNFR1-mediated bioactivity in vitro and attenuates inflammatory conditions in vivo. In this study, we aimed to improve the TNFR1-selectivity of mutTNF This was achieved by constructing a phage library displaying mutTNF-based variants, in which the amino acid residues at the predicted receptor binding sites were substituted to other amino acids. From this mutant TNF library, 20 candidate TNFR1-selective antagonists were isolated. Like mutTNF, all 20 candidates were found to have an inhibitory effect on TNFR1-mediated bioactivity. However, one of the mutants, N7, displayed significantly more than 40-fold greater TNFR1-selectivty than mutTNF. Therefore, N7 could be a promising anti-autoimmune agent that does not interfere with TNFR2-mediated signaling pathways.
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Receptores Tipo I de Factores de Necrosis Tumoral/antagonistas & inhibidores , Receptores Tipo I de Factores de Necrosis Tumoral/genética , Factores de Necrosis Tumoral/genética , Factores de Necrosis Tumoral/farmacología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Variación Genética , Humanos , Mutación , Biblioteca de Péptidos , Receptores Tipo II del Factor de Necrosis Tumoral/efectos de los fármacos , Resonancia por Plasmón de SuperficieRESUMEN
The photoluminescence spectra from a quantum-dot exciton weakly-coupled to a planar photonic-crystal cavity is experimentally investigated by temperature tuning. Significant resonance shifts of the cavity mode are observed as the cavity mode spectrally approaches that of the exciton mode, showing the appearance of cavity-to-exciton attraction or mode pulling. Cavity-mode spectral shifts are also found theoretically using a master equation model that includes incoherent pump processes for the coupled exciton and cavity, pure dephasing, and allows for photon emission via radiation modes and the leaky cavity mode. Both experiments and theory show clear cavity mode spectral shifts in the photoluminescence spectra, when certain coupling parameters are met. However, discrepancies between the experimental data and theory, including more pronounced spectral shifts in the measurements, indicate that other unknown mode-pulling effects may also be occurring.
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Cross sections for the 3He(e,e' pn)1H reaction were measured for the first time at energy transfers of 220 and 270 MeV for several momentum transfers ranging from 300 to 450 MeV/c. Cross sections are presented as a function of the momentum of the recoil proton and the momentum transfer. Continuum Faddeev calculations using the Argonne V18 and Bonn-B nucleon-nucleon potentials overestimate the measured cross sections by a factor 5 at low recoil proton momentum with the discrepancy becoming smaller at higher recoil proton momentum.
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Here we report the first demonstration of entanglement distribution over a record distance of 200 km which is of sufficient fidelity to realize secure communication. In contrast to previous entanglement distribution schemes, we use detection elements based on practical avalanche photodiodes (APDs) operating in a self-differencing mode. These APDs are low-cost, compact and easy to operate requiring only electrical cooling to achieve high single photon detection efficiency. The self-differencing APDs in combination with a reliable parametric down-conversion source demonstrate that entanglement distribution over ultra-long distances has become both possible and practical. Consequently the outlook is extremely promising for real world entanglement-based communication between distantly separated parties.