Expression and function of CCN2-derived circRNAs in chondrocytes

Cellular communication network factor 2 (CCN2) molecules promote endochondral ossification and articular cartilage regeneration, and circular RNAs (circRNAs), which arise from various genes and regulate gene expression by adsorbing miRNAs, are known to be synthesized from CCN2 in human vascular endothelial cells and other types of cells. However, in chondrocytes, not only the function but also the presence of CCN2-derived circRNA remains completely unknown. In the present study, we investigated the expression and function of CCN2-derived circRNAs in chondrocytes. Amplicons smaller than those from known CCN2-derived circRNAs were observed using RT-PCR analysis that could specifically amplify CCN2-derived circRNAs in human chondrocytic HCS-2/8 cells. The nucleotide sequences of the PCR products indicated novel circRNAs in the HCS-2/8 cells that were different from known CCN2-derived circRNAs. Moreover, the expression of several Ccn2-derived circRNAs in murine chondroblastic ATDC5 cells was confirmed and observed to change alongside chondrocytic differentiation. Next, one of these circRNAs was knocked down in HCS-2/8 cells to investigate the function of the human CCN2-derived circRNA. As a result, CCN2-derived circRNA knockdown significantly reduced the expression of aggrecan mRNA and proteoglycan synthesis. Our data suggest that CCN2-derived circRNAs are expressed in chondrocytes and play a role in chondrogenic differentiation

Regulation of cellular communication network factor 2 (CCN2) in breast cancer cells via the cell-type dependent interplay between CCN2 and glycolysis

Objectives: Anti-osteoclastic treatments for breast cancer occasionally cause medication-related osteonecrosis of the jaw. Moreover, elevated glycolytic activity, which is known as the Warburg effect, is usually observed in these breast cancer cells. Previously, we found that cellular communication network factor 2 (CCN2) production and glycolysis enhanced each other in chondrocytes. Here, we evaluated the interplay between CCN2 and glycolysis in breast cancer cells, as we suspected a possible involvement of CCN2 in the Warburg effect in highly invasive breast cancer cells. Methods: Two human breast cancer cell lines with a distinct phenotype were used. Glycolysis was inhibited by using 2 distinct compounds, and gene silencing was performed using siRNA. Glycolysis and the expression of relevant genes were monitored via colorimetric assays and quantitative RT-PCR, respectively. Results: Although CCN2 expression was almost completely silenced when treating invasive breast cancer cells with a siRNA cocktail against CCN2, glycolytic activity was not affected. Notably, the expression of glycolytic enzyme genes, which was repressed by CCN2 deficiency in chondrocytes, tended to increase upon CCN2 silencing in breast cancer cells. Inhibition of glycolysis, which resulted in the repression of CCN2 expression in chondrocytic cells, did not alter or strongly enhanced CCN2 expression in the invasive and non-invasive breast cancer cells, respectively. Conclusions: High CCN2 expression levels play a critical role in the invasion and metastasis of breast cancer. Thus, a collapse in the intrinsic repressive machinery of CCN2 due to glycolysis may induce the acquisition of an invasive phenotype in breast cancer cells

Neuroprotective response after photodynamic therapy: Role of vascular endothelial growth factor

Background: Anti-vascular endothelial growth factor (VEGF) drugs and/or photodynamic therapy (PDT) constitute current treatments targeting pathological vascular tissues in tumors and age-related macular degeneration. Concern that PDT might induce VEGF and exacerbate the disease has led us to current practice of using anti-VEGF drugs with PDT simultaneously. However, the underlying molecular mechanisms of these therapies are not well understood. Methods: We assessed VEGF levels after PDT of normal mouse retinal tissue, using a laser duration that did not cause obvious tissue damage. To determine the role of PDT-induced VEGF and its downstream signaling, we intravitreally injected a VEGF inhibitor, VEGFR1 Fc, or a PI3K/Akt inhibitor, LY294002, immediately after PDT. Then, histological and biochemical changes of the retinal tissue were analyzed by immunohistochemistry and immunoblot analyses, respectively. Results: At both the mRNA and protein levels, VEGF was upregulated immediately and transiently after PDT. VEGF suppression after PDT resulted in apoptotic destruction of the photoreceptor cell layer in only the irradiated area during PDT. Under these conditions, activation of the anti-apoptotic molecule Akt was suppressed in the irradiated area, and levels of the pro-apoptotic protein BAX were increased. Intravitreal injection of a PI3K/Akt inhibitor immediately after PDT increased BAX levels and photoreceptor cell apoptosis. Conclusion: Cytotoxic stress caused by PDT, at levels that do not cause overt tissue damage, induces VEGF and activates Akt to rescue the neural tissue, suppressing BAX. Thus, the immediate and transient induction of VEGF after PDT is neuroprotective

Long-term outcomes for Asian patients with X-linked hypophosphataemia : rationale and design of the SUNFLOWER longitudinal, observational cohort study

Introduction X-linked hypophosphataemic rickets/osteomalacia (XLH) is a chronic, debilitating genetic disease characterised by skeletal abnormalities and growth disorder. The burden of XLH begins in childhood and continues throughout life. Conventional medical therapy with phosphate, active vitamin D and surgery do not address the underlying pathophysiology of the disease. While treatment during childhood may improve bone deformity and growth retardation, a large proportion of adult patients still fail to reach normal stature. Furthermore, adult patients with XLH report comorbidities associated with unresolved childhood disease, as well as newly developed disease-related complications and significantly impaired quality of life (QOL). Despite the multiple negative aspects of XLH, Asian consensus statements for diagnosis and management are lacking. Methods and analysis The Study of longitUdinal observatioN For patients with X-Linked hypOphosphataemic rickets/osteomalacia in collaboration With Asian partnERs study is a longitudinal observational cohort study of patients with XLH, designed to determine the medical characteristics and burdens (physical, emotional and financial) of this progressive disease and to evaluate the impact of treatment (including the use of burosumab) on clinical outcomes. The study was initiated in April 2018, and registration will remain open until 30 April 2022. The sample size planned for analyses is 160 patients, consisting of 100 patients in Japan and 60 patients in Korea. Up to 5 years of observation are planned per patient, from enrolment through to April 2023. Prospective and retrospective data will be collected to evaluate variables, including height/growth, rickets severity score, QOL, motor function and biomarkers for phosphate metabolism and bone turnover. Ethics and dissemination Ethics approval was obtained from the Ethics Committee of Osaka University, the Ethics Committee of Kyowa Kirin Co and by the Ethics Committee of each participating medical institution. Two interim analyses and associated publications are planned using retrospective and enrolment data at year 1 and results at year 3

Macrophage centripetal migration drives spontaneous healing process after spinal cord injury.

Traumatic spinal cord injury (SCI) brings numerous inflammatory cells, including macrophages, from the circulating blood to lesions, but pathophysiological impact resulting from spatiotemporal dynamics of macrophages is unknown. Here, we show that macrophages centripetally migrate toward the lesion epicenter after infiltrating into the wide range of spinal cord, depending on the gradient of chemoattractant C5a. However, macrophages lacking interferon regulatory factor 8 (IRF8) cannot migrate toward the epicenter and remain widely scattered in the injured cord with profound axonal loss and little remyelination, resulting in a poor functional outcome after SCI. Time-lapse imaging and P2X/YRs blockade revealed that macrophage migration via IRF8 was caused by purinergic receptors involved in the C5a-directed migration. Conversely, pharmacological promotion of IRF8 activation facilitated macrophage centripetal movement, thereby improving the SCI recovery. Our findings reveal the importance of macrophage centripetal migration via IRF8, providing a novel therapeutic target for central nervous system injury

A Novel CLEIA for FGF23

Introduction: Measurement of fibroblast growth factor 23 (FGF23) has been reported to be clinically useful for the differential diagnosis of chronic hypophosphatemia. However, assays for research use only are available in Japan. Thus, the objective of this study was to examine the clinical utility of a novel and automated chemiluminescent enzyme immunoassay for the measurement of FGF23. Materials and Methods: Participants were recruited from July 2015 to January 2017 at six facilities in Japan. Thirty-eight patients with X-linked hypophosphatemic rickets (XLH; 15 males, 23 females, age 0–66 years), five patients with tumour-induced osteomalacia (TIO; 3 males, 2 females, age 60–73 years), and twenty-two patients with hypophosphatemia (11 males, 11 females, age 1–75 years) caused due to other factors participated in this study. Results: With the clinical cut-off value of FGF23 at 30.0 pg/mL indicated in the Diagnostic Guideline of Rickets/Osteomalacia in Japan, the sensitivity and specificity of FGF23-related hypophosphatemic rickets/osteomalacia without vitamin D deficiency (disease group-1) were 100% and 81.8%, respectively, which distinguished it from non-FGF23-related hypophosphatemia (disease group-2). Furthermore, the diagnostic sensitivity of FGF23-related hypophosphatemia with vitamin D deficiency remained at 100%. Among the four patients with FGF23 levels ≥ 30.0 pg/mL in disease group-2, two patients with relatively higher FGF23 values were suspected to have genuine FGF23-related hypophosphatemia, due to the ectopic production of FGF23 in pulmonary and prostate small cell carcinomas. Conclusion: The novel FGF23 assay tested in this study is useful for the differential diagnosis of hypophosphatemic rickets/osteomalacia in a clinical setting

Heusler alloys for spintronic devices: review on recent development and future perspectives

Heusler alloys are theoretically predicted to become half-metals at room temperature (RT). The advantages of using these alloys are good lattice matching with major substrates, high Curie temperature above RT and intermetallic controllability for spin density of states at the Fermi energy level. The alloys are categorised into half- and full-Heusler alloys depending upon the crystalline structures, each being discussed both experimentally and theoretically in Section 2. Fundamental properties of ferromagnetic Heusler alloys are described in Section 3. Both structural and magnetic characterisations on an atomic scale are typically carried out in order to prove the half-metallicity at RT as described in Section 4. Atomic ordering in the Heusler-alloy films is directly observed by X-ray diffraction and is also indirectly probed via the temperature dependence of electrical resistivity. Element specific magnetic moments and spin polarisation of the Heusler alloy films are directly measured using X-ray magnetic circular dichroism and Andreev reflection, respectively. By employing these ferromagnetic alloy films in a spintronic device, efficient spin injection into a non-magnetic material and large magnetoresistance are discussed in Section 5. Fundamental properties of antiferromagnetic Heusler alloys are described in Section 6. Both structural and magnetic characterisations on an atomic scale are shown in Section 7. Atomic ordering in the Heusler-alloy films is indirectly measured by the temperature dependence of electrical resistivity. Antiferromagnetic configurations are directly imaged by X-ray magnetic linear dichroism and polarised neutron reflection. Section 8 explains applications of the antiferromagnetic Heusler-alloy films. The other non-magnetic Heusler alloys are listed in Section 9. A brief summary is provided at the end of this review

Sutura do menisco pela técnica all-inside com o dispositivo Fast-Fix

OBJECTIVE:The objective of this study is to evaluate the results and effectiveness of the technique of meniscal repair type all-inside using Fast-Fix device.METHODS:A retrospective cohort study evaluating 22 patients with meniscal surgery between January 2004 and December 2010 underwent meniscal repair technique for all-inside with the Fast-Fix device with or without ACL reconstruction. Function and quality of life outcomes were chosen by the IKDC and Lysholm score, before and postoperatively, and reoperation rates, relying to the time of final follow-up. Statistical analysis was performed using the Student's t test.RESULTS:The mean follow-up was 59 months (16-84). The Lysholm score showed 72% (16 patients) of excellent and good results (84-100 points), 27% (6 patients) fair (65-83 points) and no cases classified as poor (<64 points). According to the IKDC: 81% (18 patients) of excellent and good results (75-100 points), 18% of cases regular (50-75 points) and no patient had poor results (<50 points). There were no failures or complications.CONCLUSION:The technique of meniscal repair type all-inside using the Fast-Fix device is safe and effective for the treatment of meniscal lesions in the red zone or red-white with or without simultaneous ACL reconstruction, with good and excellent results in most patients Level 4 Study.OBJETIVO:Avaliar os resultados e a eficácia da técnica de reparo meniscal tipo all-inside com o uso do dispositivo FasT-Fix.MÉTODOS:Estudo de coorte retrospectivo com avaliação de 22 pacientes com lesão meniscal operados entre janeiro de 2004 e dezembro de 2010, submetidos ao reparo meniscal pela técnica all-inside com o dispositivo FasT-Fix e associados ou não à reconstrução do LCA. Função e qualidade de vida foram os desfechos escolhidos por meio dos questionários de Lysholm e IKDC, pré e pós-operatoriamente, além das taxas de reoperação, relevando-se o tempo de seguimento final. A análise estatística foi feita com o uso do teste t de Student.RESULTADOS:O tempo médio de seguimento foi de 59 meses (16-84). O escore de Lysholm apresentou 73% (16 pacientes) de excelentes e bons resultados (84-100 pontos), 27% (seis pacientes) regulares (65-83 pontos) e nenhum caso classificado como ruim (< 64 pontos). Segundo o IKDC: 82% (18 pacientes) de excelentes e bons resultados (75-100 pontos); 18% de casos regulares (50-75 pontos) e nenhum paciente obteve resultados ruins (< 50 pontos). Não ocorreram falhas ou complicações.CONCLUSÃO:A técnica de reparo meniscal tipo all-inside com o uso do dispositivo FasT-Fix, nos pacientes avaliados, se mostrou eficaz e segura para o tratamento das lesões de menisco na zona vermelha ou zona vermelho-branca associada ou não à reconstrução simultânea do LCA e apresentou resultados bons e excelentes na maioria dos pacientes.Universidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Department of Orthopedics and TraumatologyUNIFESP-EPM Department of Orthopedics and TraumatologyUNIFESP, EPM, Department of Orthopedics and TraumatologyUNIFESP, EPM Department of Orthopedics and TraumatologySciEL

Efficacy of Platelet-Rich Plasma for Bone Fusion in Transforaminal Lumbar Interbody Fusion

Study DesignRetrospective case series.PurposeTo examine the efficacy of platelet-rich plasma (PRP) for bone fusion in transforaminal lumbar interbody fusion (TLIF) using local bone grafting.Overview of LiteratureSeveral authors have reported the efficacy of PRP for bone union in animal models. However, the use of PRP for bone fusion in TLIF surgery has not been fully explored.MethodsTwenty patients underwent single-level TLIF surgery because of L4 spondylolisthesis. An interbody fusion cage and local bone were used in nine patients (control group) and an interbody fusion cage, local bone, and PRP were used in 11 patients (PRP group). PRP was prepared from the patients' blood samples (400 mL) immediately before surgery. The duration of bone union and postoperative bone fusion rate were assessed using plain radiography at every 3 months postoperatively and computed tomography at 12 or 24 months postoperatively, respectively. Lower back pain, leg pain, and leg numbness were evaluated using the visual analog scale preoperatively and at 3, 6, 12, and 24 months postoperatively.ResultsThe platelet count was 8.7 times higher in PRP than in blood. The bone union rate was significantly superior in the PRP group than in the control group (91% and 77%, respectively; p=0.035), whereas the average duration of bone union was not significantly different between the groups (7.7±0.74 and 10.0±2.00 months, respectively; p=0.131). There was no significant difference in lower back pain, leg pain, and leg numbness in both groups during follow-up (p>0.05).ConclusionsOur study suggests that the use of PRP in TLIF surgery increases bone fusion rate