ABSTRACT
BACKGROUND: Rubella virus-induced granulomas have been described in patients with various inborn errors of immunity. Most defects impair T-cell immunity, suggesting a critical role of T cells in rubella elimination. However, the molecular mechanism of virus control remains elusive. OBJECTIVE: This study sought to understand the defective effector mechanism allowing rubella vaccine virus persistence in granulomas. METHODS: Starting from an index case with Griscelli syndrome type 2 and rubella skin granulomas, this study combined an international survey with a literature search to identify patients with cytotoxicity defects and granuloma. The investigators performed rubella virus immunohistochemistry and PCR and T-cell migration assays. RESULTS: This study identified 21 patients with various genetically confirmed cytotoxicity defects, who presented with skin and visceral granulomas. Rubella virus was demonstrated in all 12 accessible biopsies. Granuloma onset was typically before 2 years of age and lesions persisted from months to years. Granulomas were particularly frequent in MUNC13-4 and RAB27A deficiency, where 50% of patients at risk were affected. Although these proteins have also been implicated in lymphocyte migration, 3-dimensional migration assays revealed no evidence of impaired migration of patient T cells. Notably, patients showed no evidence of reduced control of concomitantly given measles, mumps, or varicella live-attenuated vaccine or severe infections with other viruses. CONCLUSIONS: This study identified lymphocyte cytotoxicity as a key effector mechanism for control of rubella vaccine virus, without evidence for its need in control of live measles, mumps, or varicella vaccines. Rubella vaccine-induced granulomas are a novel phenotype with incomplete penetrance of genetic disorders of cytotoxicity.
Subject(s)
Granuloma/etiology , Rubella Vaccine/adverse effects , T-Lymphocytes/immunology , Child , Child, Preschool , Female , Granuloma/genetics , Granuloma/immunology , Granuloma/virology , Humans , Infant , Phenotype , Rubella/genetics , Rubella/immunology , Rubella/virology , Skin/immunology , Skin/virologySubject(s)
Glaucoma , Optic Disk , Humans , Optic Disk/blood supply , Retina , Glaucoma/etiology , Choroid/blood supplyABSTRACT
Chronic lung allograft dysfunction, the major cause of death following lung transplantation, usually manifests as irreversible airflow obstruction associated with obliterative bronchiolitis (OB), a lesion characterized by chronic inflammation, lymphoid neogenesis, fibroproliferation and small airway obliteration. Spleen tyrosine kinase (Syk), a tyrosine kinase that regulates B cell function and innate immunity, has been implicated in the pathogenesis of chronic inflammation and tissue repair. This study evaluated the role of Syk in development of OB, using an intrapulmonary tracheal transplant model of OB with the conditional Syk-knockout Syk(flox/flox) //rosa26-CreER(T2) mice and a Syk-selective inhibitor, GSK2230413. BALB/c trachea allografts were transplanted into Syk-knockout (Syk(del/del) ) mice or wild-type C57BL/6 recipients treated with GSK2230413. At day 28, histological analysis revealed that in the Syk(del/del) and GSK2230413-treated C57BL/6 recipients, the graft lumen remained open compared with allografts transplanted into Syk-expressing (Syk(flox/flox) ) and placebo control-treated C57BL/6 recipients. Immunofluorescence showed lymphoid neogenesis with distinct B and T cell zones in control mice. In contrast, lymphoid neogenesis was absent and few B or T cells were found in Syk(del/del) and GSK2230413-treated mice. These observations suggest that inhibition of Syk may be a potential therapeutic strategy for the management of OB following lung transplantation.
Subject(s)
Bronchiolitis Obliterans/etiology , Graft Rejection/immunology , Intracellular Signaling Peptides and Proteins/physiology , Lung Transplantation/adverse effects , Lymphoid Tissue/immunology , Protein-Tyrosine Kinases/physiology , Spleen/enzymology , Animals , B-Lymphocytes/immunology , Blotting, Western , Cells, Cultured , Fibrosis/etiology , Fibrosis/pathology , Flow Cytometry , Graft Survival/immunology , Immunity, Innate/immunology , Immunoenzyme Techniques , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , Postoperative Complications , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Spleen/immunology , Syk Kinase , T-Lymphocytes/immunology , Trachea/transplantation , Transplantation, HomologousABSTRACT
Donor-specific HLA antibodies (DSA) have an adverse effect on short-term and long-term lung transplant outcomes. We implemented a perioperative strategy to treat DSA-positive recipients, leading to equivalent rejection and graft survival outcomes. Pretransplant DSA were identified to HLA-A, B, C, DR and DQ antigens. DSA-positive patients were transplanted if panel reactive antibody (PRA) ≥30% or medically urgent and desensitized with perioperative plasma exchange, intravenous immune globulin, antithymocyte globulin (ATG), and mycophenolic acid (MPA). PRA-positive/DSA-negative recipients received MPA. Unsensitized patients received routine cyclosporine, azathioprine and prednisone without ATG. From 2008-2011, 340 lung-only first transplants were performed: 53 DSA-positive, 93 PRA-positive/DSA-negative and 194 unsensitized. Thirty-day survival was 96 %/99%/96% in the three groups, respectively. One-year graft survival was 89%/88%/86% (p = 0.47). DSA-positive and PRA-positive/DSA-negative patients were less likely to experience any ≥ grade 2 acute rejection (9% and 9% vs. 18% unsensitized p = 0.04). Maximum predicted forced expiratory volume (1 s) (81%/74%/76%, p = NS) and predicted forced vital capacity (81%/77%/78%, respectively, p = NS) were equivalent between groups. With the application of this perioperative treatment protocol, lung transplantation can be safely performed in DSA/PRA-positive patients, with similar outcomes to unsensitized recipients.
Subject(s)
Desensitization, Immunologic/methods , Graft Survival/physiology , Lung Transplantation/mortality , Lung/physiology , Perioperative Care/methods , Transplant Recipients , Adult , Aged , Antilymphocyte Serum/therapeutic use , Canada , Cohort Studies , Female , Follow-Up Studies , Forced Expiratory Volume/physiology , Humans , Immunoglobulins, Intravenous/therapeutic use , Lung/surgery , Male , Middle Aged , Mycophenolic Acid/therapeutic use , Plasma Exchange , Retrospective Studies , Treatment Outcome , Vital Capacity/physiologyABSTRACT
BACKGROUND: Syk, an immune regulatory tyrosine kinase, plays a role in inflammatory disease processes. We recently reported a role for epithelial expression of Syk in the airways hyper-responsiveness in response to air pollution in a mouse model of asthma. The aim of this study was to further investigate the role of Syk in airway contractility in response to methacholine (MCh) and particulate matter (PM) air pollutants, in the absence of underlying inflammation. METHODS: We used Syk(flox/flox) //rosa26CreER(T) (2) conditional Syk knockout mice to evaluate respiratory mechanics and MCh responsiveness following PM exposure in vivo using the ventilator-based flexiVent system. RESULTS: While total and differential cell counts in bronchoalveolar lavage fluid were similar between the Syk(flox/flox) and Syk(del/del) mice, central airways respiratory resistance (RN ) to MCh was significantly augmented following PM exposure between Syk-intact (Syk(flox/flox) ) and Syk-deficient (Syk(del/del) ) mice (RN (max) : 2.06 ± 0.29 vs. 1.29 ± 0.10, respectively; p < 0.05, n = 8-10/group). We employed live videomicroscopy to investigate changes in airway luminal diameter using ex vivo lung slices, which were devoid of circulating leukocytes. MCh reduced the airway luminal area of Syk(flox/flox) mice to 81.1 ± 1.4% of baseline, which was virtually abrogated in Syk(del/del) mice (luminal area = 93.2 ± 0.5%, n = 5/group, p < 0.05). In response to PM exposure, Syk(flox/flox) airways contracted to 73.8 ± 2.7% of baseline luminal diameter, whereas Syk(del/del) airways exhibited minimal contractility to PM and MCh (90.0 ± 1.3% of baseline, n = 5/group, p < 0.05). CONCLUSIONS: These observations suggest that Syk mediates airway contractility in the normal and allergic airways, independent of its role and function in leukocytes, and supports a paracrine role for airway epithelial Syk in modulating airway smooth muscle activity.
Subject(s)
Intracellular Signaling Peptides and Proteins/genetics , Leukocytes/metabolism , Protein-Tyrosine Kinases/genetics , Respiratory Hypersensitivity/genetics , Respiratory Hypersensitivity/physiopathology , Air Pollutants/adverse effects , Airway Resistance/genetics , Airway Resistance/immunology , Animals , Bronchial Provocation Tests , Bronchoalveolar Lavage Fluid/cytology , Bronchoalveolar Lavage Fluid/immunology , Cell Count , Disease Models, Animal , Gene Deletion , Intracellular Signaling Peptides and Proteins/metabolism , Leukocytes/immunology , Methacholine Chloride/administration & dosage , Mice , Mice, Knockout , Particulate Matter/adverse effects , Phenotype , Protein-Tyrosine Kinases/metabolism , Respiratory Hypersensitivity/immunology , Respiratory Hypersensitivity/metabolism , Syk KinaseABSTRACT
Chronic lung allograft dysfunction (CLAD) is the leading cause of mortality following lung transplantation. We conducted a retrospective cohort study including 397 bilateral lung recipients transplanted in from 1996 to 2009 to determine the association between ambient air pollution, CLAD and mortality. Pollution exposure was assessed using satellite-based estimates of nitrogen dioxide, distance to major roadway and total length of roadways around a patient's home. Cumulative exposures to ozone and particulate matter were estimated from concentrations measured at fixed-site stations near patients' homes using inverse distance weighted interpolation. Cox proportional hazards models were used to estimate the associations of CLAD with air pollution exposure, adjusting for various individual and neighborhood characteristics. During the follow-up, 185 patients developed CLAD (47%) and 101 patients died (25%). Fifty-four deaths (53%) were due to CLAD. We observed an association between CLAD development and road density within 200 m of a patient's home (HR 1.30 [95% CI 1.07-1.58]). Although based on a subgroup of 14 patients, living within 100 m of a highway was associated with a high risk for developing CLAD (HR 4.91 [95% CI 2.22, 10.87]). These data suggest that exposure to traffic-related air pollution is associated with development of CLAD among lung transplant recipients.
Subject(s)
Air Pollution/adverse effects , Graft Rejection/etiology , Lung Diseases/surgery , Lung Transplantation , Particulate Matter/adverse effects , Postoperative Complications , Adult , Aged , Allografts , Female , Follow-Up Studies , Graft Rejection/diagnosis , Graft Survival , Humans , Male , Middle Aged , Prognosis , Retrospective Studies , Risk FactorsABSTRACT
BACKGROUND: Malignant pleural mesothelioma (MPM) is a lethal neoplasm exhibiting resistance to most treatment regimens and requires effective therapeutic options. Though an effective strategy in many cancer, targeted therapy is relatively unexplored in MPM because the therapeutically important oncogenic pathways and networks in MPM are largely unknown. MATERIALS AND METHODS: We carried out gene expression microarray profiling of 53 surgically resected MPMs tumors along with paired normal tissue. We also carried out whole transcriptomic sequence (RNA-seq) analysis on eight tumor specimens. Taqman-based quantitative Reverse-transcription polymerase chain reaction (qRT-PCR), western analysis and immunohistochemistry (IHC) analysis of mitotic arrest deficient-like 1 (MAD2L1) was carried out on tissue specimens. Cell viability assays of MPM cell lines were carried out to assess sensitivity to specific small molecule inhibitors. RESULTS: Bioinformatics analysis of the microarray data followed by pathway analysis revealed that the mitotic spindle assembly checkpoint (MSAC) pathway was most significantly altered in MPM tumors with upregulation of 18 component genes, including MAD2L1 gene. We validated the microarray data for MAD2L1 expression using quantitative qRT-PCR and western blot analysis on tissue lysates. Additionally, we analyzed expression of the MAD2L1 protein by IHC using an independent tissue microarray set of 80 MPM tissue samples. Robust clustering of gene expression data revealed three novel subgroups of tumors, with unique expression profiles, and showed differential expression of MSAC pathway genes. Network analysis of the microarray data showed the cytoskeleton/spindle microtubules network was the second-most significantly affected network. We also demonstrate that a nontaxane small molecule inhibitor, epothilone B, targeting the microtubules have great efficacy in decreasing viability of 14 MPM cell lines. CONCLUSIONS: Overall, our findings show that MPM tumors have significant deregulation of the MSAC pathway and the microtubule network, it can be classified into three novel molecular subgroups of potential therapeutic importance and epothilone B is a promising therapeutic agent for MPM.
Subject(s)
Lung Neoplasms/genetics , M Phase Cell Cycle Checkpoints/genetics , Mesothelioma/genetics , Microtubules/pathology , Pleural Neoplasms/genetics , Adaptor Proteins, Signal Transducing/genetics , Adaptor Proteins, Signal Transducing/metabolism , Antineoplastic Agents/pharmacology , Blotting, Western , Cell Cycle Proteins/genetics , Cell Cycle Proteins/metabolism , Cell Line, Tumor , Cluster Analysis , DNA Mutational Analysis , Epothilones/pharmacology , Gene Expression Profiling , Humans , Immunohistochemistry , Mesothelioma, Malignant , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Oligonucleotide Array Sequence Analysis , Pleural Neoplasms/pathology , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Tissue Array Analysis , Transcriptome , Tubulin Modulators/pharmacologyABSTRACT
We propose and demonstrate an orthogonal frequency division multiplexing (OFDM) radio-frequency (RF) power-fading circumvention scheme for long-reach wavelength-division-multiplexed passive-optical-network (LR-WDM-PON); hence the same capacity of 40 Gb/s can be provided to all the optical-networking-units (ONUs) in the LR-WDM-PON. Numerical analysis and proof-of-concept experiment are performed.
ABSTRACT
We propose and demonstrate the feasibility of using all-optical orthogonal frequency division multiplexing (AO-OFDM) for the convergent optical wired and wireless access networks. AO-OFDM relies on all-optically generated orthogonal subcarriers; hence, high data rate (> 100 Gb/s) can be easily achieved without hitting the speed limit of electronic digital-to-analog and analog-to-digital converters (DAC/ADC). A proof-of-concept convergent access network using AO-OFDM super-channel (SC) is demonstrated supporting 40 - 100 Gb/s wired and gigabit/s 100 GHz millimeter-wave (MMW) ROF transmissions.
ABSTRACT
We propose and experimentally demonstrate a white-light phosphor-LED visible light communication (VLC) system with an adaptive 84.44 to 190 Mbit/s 16 quadrature-amplitude-modulation (QAM) orthogonal-frequency-division-multiplexing (OFDM) signal utilizing bit-loading method. Here, the optimal analogy pre-equalization design is performed at LED transmitter (Tx) side and no blue filter is used at the Rx side. Hence, the ~1 MHz modulation bandwidth of phosphor-LED could be extended to 30 MHz. In addition, the measured bit error rates (BERs) of < 3.8 × 10(-3) [forward error correction (FEC) threshold] at different measured data rates can be achieved at practical transmission distances of 0.75 to 2 m.
ABSTRACT
In this work, a scalable and continuous upgradable convergent optical access network is proposed. By using a multi-wavelength coherent comb source and a programmable waveshaper at the central office (CO), optical millimeter-wave (mm-wave) signals of different frequencies (from baseband to > 100 GHz) can be generated. Hence, it provides a scalable and continuous upgradable solution for end-user who needs 60 GHz wireless services now and > 100 GHz wireless services in the future. During the upgrade, user only needs to upgrade their optical networking unit (ONU). A programmable waveshaper is used to select the suitable optical tones with wavelength separation equals to the desired mm-wave frequency; while the CO remains intact. The centralized characteristics of the proposed system can easily add any new service and end-user. The centralized control of the wavelength makes the system more stable. Wired data rate of 17.45 Gb/s and w-band wireless data rate up to 3.36 Gb/s were demonstrated after transmission over 40 km of single-mode fiber (SMF).
ABSTRACT
We demonstrate experimentally Manchester (MC) coding based W-band (75 - 110 GHz) radio-over-fiber (ROF) system to reduce the low-frequency-components (LFCs) signal distortion generated by two independent low-cost lasers using spectral shaping. Hence, a low-cost and higher performance W-band ROF system is achieved. In this system, direct-beating of two independent low-cost CW lasers without frequency tracking circuit (FTC) is used to generate the millimeter-wave. Approaches, such as delayed self-heterodyne interferometer and heterodyne beating are performed to characterize the optical-beating-interference sub-terahertz signal (OBIS). Furthermore, W-band ROF systems using MC coding and NRZ-OOK are compared and discussed.
Subject(s)
Fiber Optic Technology/instrumentation , Lasers , Telecommunications/instrumentation , Wireless Technology/instrumentation , Equipment DesignABSTRACT
BACKGROUND: The inflammatory immune response associated with allergic airway inflammation in asthma involves T helper type 2 (Th2) immunity. Given the data that a newly described late activator antigen-presenting cell (LAPC) population promotes Th2 immunity in viral infections, we undertook studies to investigate whether LAPCs have a pathogenic role in allergic airway inflammation. METHODS: We employed acute ovalbumin (OVA) and house dust mite (HDM) sensitization and challenge models to establish allergic airway inflammation in mice, followed by the analysis of lungs and draining lymph node (DLN) cell infiltrates, immunoglobulin E (IgE) production, and airway hyper-responsiveness (AHR). We tested whether adoptive transfer of LAPCs isolated from mice with established allergic airway inflammation augments the development of sensitization in naïve mice. RESULTS: We provide evidence that in both OVA and HDM mouse models of allergic inflammation, LAPCs accumulate in the lungs and draining lymph nodes (DLNs), concomitant with the onset of lung pathology, allergen-specific IgE production, eosinophilia, and Th2 cytokine production. Adoptive transfer experiments using OVA-activated LAPCs reveal exacerbation of disease pathology with an increase in lung inflammatory cells, eosinophilia, circulating IgE, Th2 cytokine production, and a worsening of AHR. OVA-activated LAPCs preferentially increased GATA-3 induction in naïve CD4(+) T cells. CONCLUSIONS: Together, these data suggest an important role for LAPCs in polarizing the Th2 response in mouse models of allergic airway inflammation.
Subject(s)
Antigen-Presenting Cells/immunology , Asthma/immunology , Hypersensitivity/immunology , Inflammation/immunology , Lymphocyte Activation/immunology , Adoptive Transfer , Animals , Disease Models, Animal , Female , Flow Cytometry , Immunoglobulin E/immunology , Mice , Mice, Inbred BALB C , Ovalbumin/immunology , Pyroglyphidae/immunology , Th2 Cells/immunologyABSTRACT
In this work, we propose and demonstrate a stable and wavelength-tunable erbium-doped fiber (EDF) ring laser. Here, a silicon-on-insulator (SOI)-based silicon-micro-ring-resonator (SMRR) is used as the wavelength selective element inside the fiber ring cavity. A uniform period grating coupler (GC) is used to couple between the SMRR and single mode fiber (SMF) and serves also as a polarization dependent element in the cavity. The output lasing wavelength of the proposed fiber laser can be tuned at a tuning step of 2 nm (defined by the free spectral range (FSR) of the SMRR) in a bandwidth of 35.2 nm (1532.00 to 1567.20 nm), which is defined by the gain of the EDF. The optical-signal-to-noise-ratio (OSNR) of each lasing wavelength is larger than 42.0 dB. In addition, the output stabilities of power and wavelength are also discussed.
Subject(s)
Lasers, Solid-State , Lasers , Silicon/chemistry , Transducers , Equipment Design , Equipment Failure Analysis , MiniaturizationABSTRACT
We have constructed a BAC framework map of the mouse genome consisting of 2,808 PCR-confirmed BAC clusters, using a previously described method. Fingerprints of BACs from selected clusters confirm the accuracy of the map. Combined with BAC fingerprint data, the framework map covers 37% of the mouse genome.
Subject(s)
Chromosome Mapping , Chromosomes, Artificial, Bacterial , Genome , Animals , Genetic Markers , MiceABSTRACT
To provide broadband services in a single and low cost perform, the convergent optical wired and wireless access network is promising. Here, we propose and demonstrate a convergent optical wired and wireless long-reach access networks based on orthogonal wavelength division multiplexing (WDM). Both the baseband signal and the radio-over-fiber (ROF) signal are multiplexed and de-multiplexed in optical domain, hence it is simple and the operation speed is not limited by the electronic bottleneck caused by the digital signal processing (DSP). Error-free de-multiplexing and down-conversion can be achieved for all the signals after 60 km (long-reach) fiber transmission. The scalability of the system for higher bit-rate (60 GHz) is also simulated and discussed.
ABSTRACT
In this work, we experimentally demonstrate a bi-directional transmission link using light emitting diode (LED) visible light communication (VLC) for both downlink and uplink paths. Time-division-duplex (TDD) is proposed and demonstrated to significantly eliminate the reflection interference in VLC. A free space bi-directional transmission of 2 m using simple on-off keying (OOK) modulation with bit error rate (BER) of < 10(-5) in both directions are achieved. Moreover, the influence of reflection interference is analyzed, showing the proposed scheme can significantly eliminate the reflection interference.
Subject(s)
Artifacts , Lighting/instrumentation , Optical Devices , Semiconductors , Signal Processing, Computer-Assisted/instrumentation , Telecommunications/instrumentation , Equipment Design , Equipment Failure AnalysisABSTRACT
Cystic lymphangioma is a rare lesion of the mediastinum. We present a patient with an antenatally detected mediastinal mass that appeared to regress during foetal life and was not demonstrated on early postnatal imaging. Acute severe respiratory distress at two months of age precipitated surgery with subsequent diagnosis of lymphangioma.
Subject(s)
Lymphangioma, Cystic/diagnosis , Mediastinal Neoplasms/diagnosis , Humans , Infant , Lymphangioma, Cystic/pathology , Lymphangioma, Cystic/surgery , Male , Mediastinal Neoplasms/pathology , Mediastinal Neoplasms/surgery , Respiratory Distress Syndrome , SternotomyABSTRACT
Over the past decade, several distinct novel renal epithelial neoplasms driven by underlying tuberous sclerosis comples ( TSC)/ mammalian target of rapamycin (MTOR) pathway mutations have been described. We report herein two distinctive TSC2 -mutated renal cell carcinomas which do not fit any previously described entity. The two renal carcinomas occurred in young patients (ages 10 and 31 y), and were characterized by highly permeative growth within the kidney with metastases to perirenal lymph nodes. The neoplastic cells were predominantly large, multinucleated giant cells having variably eosinophilic to xanthomatous cytoplasm with basophilic stippling and frequent vacuolization. While the discohesive nature of the neoplastic cells, xanthomatous cytoplasm, immunoreactivity for histiocytic markers and minimal immunoreactivity for conventional epithelial markers raised the possibility of a histiocytic neoplasm, multifocal immunoreactivity for cytokeratin 20 helped establish their epithelial nature. Despite the aggressive growth pattern of these neoplasms and lymph node metastases, mitotic figures were rare and Ki-67 indices were low (<1%). One patient with follow-up shows no evidence of disease seven years after nephrectomy with no adjuvant therapy. Next-generation sequencing demonstrated TSC2 mutations in each case. By immunohistochemistry, downstream markers of mTOR pathway activation S6K1, 4EBP1, and glycoprotein nonmetastatic melanoma protein B were all highly expressed in these neoplasms, suggesting mTOR pathway activation as the neoplastic driver. While the cytokeratin 20 immunoreactivity and focal basophilic cytoplasmic stippling suggest a relationship to eosinophilic solid and cystic renal cell carcinoma, and cytoplasmic vacuolization suggests a relationship to eosinophilic vacuolated tumor, these neoplasms appear to be distinctive given their permeative growth patterns and predominant xanthomatous giant cell morphology. Addition of cytokeratin 20 to a panel of epithelial markers helps avoid misdiagnosis in such cases.
Subject(s)
Carcinoma, Renal Cell , Kidney Neoplasms , Tuberous Sclerosis , Adolescent , Adult , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , Carcinoma, Renal Cell/pathology , Child , Female , Giant Cells/pathology , Glycoproteins , Humans , Keratin-20 , Ki-67 Antigen , Kidney Neoplasms/pathology , Male , TOR Serine-Threonine Kinases/genetics , Tuberous Sclerosis Complex 1 Protein , Young AdultABSTRACT
Long-reach passive optical network (LR-PON) is considered as a promising technology towards higher capacity and extended coverage optical system. We propose and demonstrate a LR-PON with the capability of Rayleigh backscattering (RB) noise mitigation. By using the upstream signal wavelength-transition generated by a dual-parallel Mach-Zehnder modulator (DP-MZM) based colorless optical networking unit (ONU), the spectral overlap among the upstream signal and the RB noises can be minimized. Hence, due to the achievement of effective RB mitigation, a 100 km LR-PON with a high split-ratio of 512 is demonstrated using 10 Gb/s non-return-to-zero (NRZ) downstream and 2.5 Gb/s NRZ upstream signals. Detail analysis of the wavelength-transition generation is presented.