RESUMO
The bleomycin-induced scleroderma model is a well-established and dependable method for creating a mouse model of SSc (systemic sclerosis). In the field of skin connective tissue diseases, increasing evidence from clinical and animal experiments suggests that TLRs (Toll-like receptors) play an important role in several diseases. This study aimed to determine the role of TLR7 (Toll-like receptor 7) and TLR9 (Toll-like receptor 9) in the mechanisms of immune abnormalities and fibrosis in SSc. This study used TLR7-KO mice (TLR7-knockout mice with a balb/c background) and TLR9-KO mice (TLR9-knockout mice with a balb/c background) as well as WT mice (wild-type balb/c mice). All three kinds of mice were induced by BLM (bleomycin) in a scleroderma model as the experimental group; meanwhile, WT mice treated with PBS (phosphate-buffered saline) were used as the control group. We analyzed the fibrotic phenotype and the immunological abnormality phenotype of TLR7-deficient and TLR9-deficient mice in the SSc disease model using flow cytometry, RT-PCR (reverse transcription-polymerase chain reaction), a histological examination, and IHC (immunohistochemical staining). In a mouse model of SSc disease, the deletion of TLR7 attenuated skin and lung fibrosis, while the deletion of TLR9 exacerbated skin and lung fibrosis. The deletion of TLR7 resulted in a relative decrease in the infiltration and expression of various pro-inflammatory and fibrotic cells and cytokines in the skin. On the other hand, the deletion of TLR9 resulted in a relative increase in the infiltration and expression of various pro-inflammatory and cytokine-inhibiting cells and cytokines in the skin. Under the influence of pDCs (plasmacytoid dendritic cells), the balances of Beff/Breg (IL-6 + CD19 + B cell/IL-10 + CD19 + B cell), Th17/Treg (IL-17A + CD4 + T cell/Foxp3 + CD25 + CD4 + T cell), M1/M2 (CD86 + macrophage/CD206 + macrophage), and Th1/Th2 (TNFα + CD3 + CD4 + T cell/IL-4 + CD3 + CD4 + T cell) were biased towards the suppression of inflammation and fibrosis as a result of the TLR7 deletion. Comparatively, the balance was biased towards promoting inflammation and fibrosis due to the TLR9 deletion. In the SSc model, TLR7 promoted inflammation and fibrosis progression, while TLR9 played a protective role. These results suggest that TLR7 and TLR9 play opposite roles in triggering SSc to produce immune system abnormalities and skin fibrosis.
Assuntos
Modelos Animais de Doenças , Camundongos Knockout , Escleroderma Sistêmico , Receptor 7 Toll-Like , Receptor Toll-Like 9 , Animais , Receptor 7 Toll-Like/metabolismo , Receptor 7 Toll-Like/genética , Escleroderma Sistêmico/metabolismo , Escleroderma Sistêmico/patologia , Escleroderma Sistêmico/imunologia , Escleroderma Sistêmico/genética , Receptor Toll-Like 9/metabolismo , Receptor Toll-Like 9/genética , Camundongos , Bleomicina/efeitos adversos , Camundongos Endogâmicos BALB C , Citocinas/metabolismo , Pele/patologia , Pele/metabolismo , Pele/imunologia , Fibrose , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/patologia , Fibrose Pulmonar/etiologia , Glicoproteínas de MembranaRESUMO
The emergence of tyrosine kinase inhibitors as part of a front-line treatment has greatly improved the clinical outcome of the patients with Ph+ acute lymphoblastic leukemia (ALL). However, a portion of them still become refractory to the therapy mainly through acquiring mutations in the BCR-ABL1 gene, necessitating a novel strategy to treat tyrosine kinase inhibitor (TKI)-resistant Ph+ ALL cases. In this report, we show evidence that RUNX1 transcription factor stringently controls the expression of BCR-ABL1, which can strategically be targeted by our novel RUNX inhibitor, Chb-M'. Through a series of in vitro experiments, we identified that RUNX1 binds to the promoter of BCR and directly transactivates BCR-ABL1 expression in Ph+ ALL cell lines. These cells showed significantly reduced expression of BCR-ABL1 with suppressed proliferation upon RUNX1 knockdown. Moreover, treatment with Chb-M' consistently downregulated the expression of BCR-ABL1 in these cells and this drug was highly effective even in an imatinib-resistant Ph+ ALL cell line. In good agreement with these findings, forced expression of BCR-ABL1 in these cells conferred relative resistance to Chb-M'. In addition, in vivo experiments with the Ph+ ALL patient-derived xenograft cells showed similar results. In summary, targeting RUNX1 therapeutically in Ph+ ALL cells may lead to overcoming TKI resistance through the transcriptional regulation of BCR-ABL1. Chb-M' could be a novel drug for patients with TKI-resistant refractory Ph+ ALL.
Assuntos
Subunidade alfa 2 de Fator de Ligação ao Core/metabolismo , Proteínas de Fusão bcr-abl/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Animais , Antineoplásicos/farmacologia , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/genética , Subunidade alfa 2 de Fator de Ligação ao Core/antagonistas & inibidores , Subunidade alfa 2 de Fator de Ligação ao Core/genética , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Mesilato de Imatinib/farmacologia , Camundongos , Mutação , Cromossomo Filadélfia , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Inibidores de Proteínas Quinases/farmacologiaRESUMO
Ascertaining the structure and functions of mitochondrial respiratory chain complexes is essential to understanding the biological mechanisms of energy conversion; therefore, numerous studies have examined these complexes. A fundamental part of that research involves devising a method for purifying samples with good reproducibility; the samples obtained need to be stable and their constituents need to retain the same structure and functions they possess when in mitochondrial membranes. Submitochondrial bovine heart particles were isolated using differential centrifugation to adjust to a membrane concentration of 46.0% (w/v) or 31.5% (w/v) based on weight. After 0.7% (w/v) deoxycholic acid, 0.4% (w/v) decyl maltoside, and 7.2% (w/v) potassium chloride were added to the mitochondrial membranes, those membranes were solubilized. At a membrane concentration of 46%, complex V was selectively solubilized, whereas at a concentration of 31.5% (w/v), complexes I and III were solubilized. Two steps-sucrose density gradient centrifugation and anion-exchange chromatography on a POROS HQ 20⯵m column-enabled selective purification of samples that retained their structure and functions. These two steps enabled complexes I, III, and V to be purified in two days with a high yield. Complexes I, III, and V were stabilized with n-decyl-ß-D-maltoside. A total of 200â¯mg-300â¯mg of those complexes from one bovine heart (1.1â¯kg muscle) was purified with good reproducibility, and the complexes retained the same functions they possessed while in mitochondrial membranes.
Assuntos
Complexo III da Cadeia de Transporte de Elétrons , Complexo I de Transporte de Elétrons , Mitocôndrias Cardíacas/enzimologia , Membranas Mitocondriais/enzimologia , Proteínas Mitocondriais , ATPases Mitocondriais Próton-Translocadoras , Miocárdio/enzimologia , Animais , Bovinos , Complexo I de Transporte de Elétrons/química , Complexo I de Transporte de Elétrons/isolamento & purificação , Complexo III da Cadeia de Transporte de Elétrons/química , Complexo III da Cadeia de Transporte de Elétrons/isolamento & purificação , Proteínas Mitocondriais/química , Proteínas Mitocondriais/isolamento & purificação , ATPases Mitocondriais Próton-Translocadoras/química , ATPases Mitocondriais Próton-Translocadoras/isolamento & purificação , SolubilidadeRESUMO
BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) has a dismal prognosis. Chemokines play important roles in the progression of many malignancies; however, the role of chemokine receptor expression in clinical cases of PDAC is unclear. Moreover, little is known about DARC, a decoy receptor of CXC chemokines, in the regulation of tumor progression. METHODS: Functions of chemokine receptors were evaluated using surgical specimens collected from PDAC patients, and PDAC cell lines. RESULTS: CXCR2 expression had no impacts on predicting prognosis, but low DARC expression in cancer cells was an independent risk factor for poor prognosis. In PDAC with low DARC expression, tumor sizes were larger and vascular invasion was increased. High CXCR2 expression was a significant predictor for poor prognosis, only in PDAC patients with low DARC expression. CXCR2 signaling induced STAT3 activation in PDAC, resulting in promoting cell cycle progression, inhibiting apoptosis, inducing angiogenesis, and enhancing invasiveness. DARC inhibited STAT3 activation by down-regulating CXCR2 signaling. These effects were confirmed by EMSA in vitro. DARC knockdown significantly increased cell proliferation in CFPAC-1 cells with high DARC expression, by activating STAT3. In contrast, CXCR2 knockdown inhibited the proliferative effects of IL-8 in MIA PaCa-2 cells with low DARC expression. Moreover, the inhibitory effect of CXCR2 antagonist on PDAC cell proliferation was more powerful in MIA PaCa-2 cells than CFPAC-1 cells. CONCLUSIONS: DARC expressing in cancer cells inhibits PDAC progression by suppressing STAT3 activation through the inhibition of CXCR2 signaling. Therefore, DARC is a novel prognostic predictor and a potential therapeutic target for PDAC.
Assuntos
Carcinoma Ductal Pancreático/metabolismo , Sistema do Grupo Sanguíneo Duffy/metabolismo , Neoplasias Pancreáticas/metabolismo , Receptores de Superfície Celular/metabolismo , Receptores de Interleucina-8B/antagonistas & inibidores , Idoso , Carcinoma Ductal Pancreático/imunologia , Carcinoma Ductal Pancreático/mortalidade , Carcinoma Ductal Pancreático/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Progressão da Doença , Feminino , Humanos , Interleucina-8/farmacologia , Masculino , Pessoa de Meia-Idade , Neoplasias Pancreáticas/imunologia , Neoplasias Pancreáticas/mortalidade , Neoplasias Pancreáticas/patologia , Receptores de Superfície Celular/antagonistas & inibidores , Receptores de Interleucina-8A/metabolismo , Receptores de Interleucina-8B/metabolismo , Fator de Transcrição STAT3/metabolismo , Transdução de SinaisRESUMO
Aggressive digital papillary adenocarcinoma (ADPA) is a rare sweat gland neoplasm with a high recurrence rate and metastatic potential. In this study, the authors describe a case that originally appeared to benign spiradenoma, but took an ominous course eventually resulting in the diagnosis of ADPA. A 73-year-old woman developed a gradually growing nodule on the second toe of her left foot, which she had first noticed 4 years previously. An excisional biopsy was performed followed by histological examination. The authors initially considered the tumor to be a benign spiradenoma and did not perform reexcision. However, she experienced local recurrence 24 months later, and multiple pulmonary metastasis 31 months later. On histological examination, both the primary and locally recurrent tumors were found to be composed of discrete and well-circumscribed solid nodules, lacking cystic space. All tumors (the primary tumor, locally recurrent tumor, and lung metastases) presented with a pattern of fused back-to-back tubular structures and myoepithelial differentiation confirmed by immunohistochemical examination. On the basis of these findings, the authors finally diagnosed ADPA with multiple pulmonary metastases. The patient underwent chemotherapy, but died of disease 49 months later. This case highlights the importance of high clinical suspicion of ADPA when digital lesions present.
Assuntos
Adenocarcinoma Papilar/secundário , Neoplasias Pulmonares/secundário , Neoplasias das Glândulas Sudoríparas/patologia , Dedos do Pé/patologia , Adenocarcinoma Papilar/química , Adenocarcinoma Papilar/tratamento farmacológico , Adenocarcinoma Papilar/cirurgia , Idoso , Antineoplásicos/uso terapêutico , Biópsia , Progressão da Doença , Evolução Fatal , Feminino , Humanos , Imuno-Histoquímica , Neoplasias Pulmonares/química , Neoplasias Pulmonares/tratamento farmacológico , Recidiva Local de Neoplasia , Neoplasias das Glândulas Sudoríparas/química , Neoplasias das Glândulas Sudoríparas/cirurgia , Fatores de Tempo , Dedos do Pé/cirurgia , Tomografia Computadorizada por Raios X , Resultado do TratamentoRESUMO
A 70-year-old woman was diagnosed with pancreatic head cancer with hepatic artery invasion by multi-detector computed tomography (MD-CT). After 3 courses of gemcitabine plus S-1 neoadjuvant therapy, the tumor size was not diminished; however, the tumor marker CA19-9 level was decreased to less than 90% of its initial level. Pancreaticoduodenectomy combined with hepatic artery resection was performed, and an end-to-end anastomosis was made between the common and proper hepatic artery to reconstruct the hepatic artery. The pathological examination indicated adenosquamous carcinoma, no vascular invasion, and negative margin status, and the efficacy of chemotherapy was classified as Gradeâ ¡b using Evans' classification. Usually, pancreatic head cancer with hepatic artery invasion is considered unresectable due to its high morbidity/mortality and poor prognosis. However, with the recently developed surgical strategy and appropriate therapeutic interventions, such as a combination of neoadjuvant chemotherapy and resection/reconstruction of the hepatic artery, a curative operation can be feasible for locally advanced pancreatic head cancer.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Artéria Hepática/cirurgia , Terapia Neoadjuvante , Neoplasias Pancreáticas/tratamento farmacológico , Idoso , Desoxicitidina/administração & dosagem , Desoxicitidina/análogos & derivados , Combinação de Medicamentos , Feminino , Artéria Hepática/patologia , Humanos , Ácido Oxônico/administração & dosagem , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/cirurgia , Pancreaticoduodenectomia , Tegafur/administração & dosagem , GencitabinaRESUMO
Interstitial lung disease (ILD) is recognized a prognostic factor and leading cause of death in patients with systemic sclerosis (SSc). The aim of the present study is to clarify factors at an initial visit that are associated with the deterioration of ILD in SSc patients with anti-topoisomerase I (anti-topo I) antibodies. This was a single-center, retrospective, observational study. Fifty-three consecutive SSc patients with anti-topo I antibodies were included in this study. Of the 53 patients, 43 had ILD at their initial visit, whereas 10 did not. We examined the clinical and immunological factors at an initial visit that were associated with the deterioration of ILD. The deterioration of ILD was defined as the administration of intravenous cyclophosphamide (IVCY) therapy. In this cohort, 45 (85%) patients had ILD at the time of the final observation, and only two who did not have ILD at their initial visit developed ILD during the follow-up period. Until the final observation, 26 (49%) patients received IVCY therapy for the progression of ILD. The age at onset, disease duration, SSc subtype, and skin score were similar between patients with and those without IVCY therapy. Approximately 60% (26 of 43) of patients with ILD at their initial visit received IVCY therapy. On the other hand, none of the 10 patients without ILD at their initial visit received IVCY therapy. Our multivariate analyses using Cox proportional hazards regression model revealed that the presence of ILD at an initial visit was an independent factor associated with the introduction of IVCY therapy (odds ratio, 2.8e+7 [95% confidence interval, 1.8e+17-uncalculated], p = 0.0048). Although anti-topo I antibodies are strongly associated with ILD, it was unlikely for SSc patients with anti-topo I antibodies to receive IVCY therapy when they did not have ILD at an initial visit.
Assuntos
Doenças Pulmonares Intersticiais , Escleroderma Sistêmico , Humanos , Imunossupressores , Estudos Retrospectivos , Japão/epidemiologia , Resultado do Tratamento , Doenças Pulmonares Intersticiais/diagnóstico , Doenças Pulmonares Intersticiais/tratamento farmacológico , Doenças Pulmonares Intersticiais/etiologia , Ciclofosfamida/uso terapêutico , Escleroderma Sistêmico/complicações , Escleroderma Sistêmico/tratamento farmacológico , PulmãoRESUMO
Anti-human upstream-binding factor (anti-hUBF) antibodies have been reported predominantly in patients with connective tissue diseases (CTDs); these have also been reported in patients without CTDs such as hepatocellular carcinoma. Because of the low frequency of expression and few case reports, there is no consensus on the clinical significance of these antibodies. Thus, we aimed to examine the clinical features of patients with anti-hUBF antibodies and analyzed 1042 patients with clinically suspected CTDs. The presence of anti-hUBF antibodies was screened using immunoprecipitation assays. Of the 1042 patients, 19 (1.82%) tested positive for anti-hUBF antibodies; among them, 10 (56%) were diagnosed with undifferentiated CTD (UCTD), six with systemic sclerosis (SSc) and three with other diseases. Five of the 10 patients with UCTD were referred to our hospital with suspected SSc. None of the five patients fulfilled the 2013 American College of Rheumatology/European League Against Rheumatism classification criteria, but three scored seven points, a relatively high score. Six anti-hUBF-positive patients with SSc had a significantly lower modified Rodnan skin score (mRSS) than that of anti-hUBF-negative patients with SSc (2 [0-2] vs 7 [0-49], p < 0.01). Compared with anti-topoisomerase I-positive patients, anti-hUBF-positive patients had a significantly lower mRSS (2 [0-2] vs 13 [0-42], p < 0.01) and lower incidence of scleroderma renal crisis (0 of 6 vs 8 of 184, p < 0.01). Compared with anti-centromere-positive patients, anti-hUBF-positive patients had a higher incidence of interstitial lung disease (ILD), but the difference was not statistically significant (4 of 6 vs 19 of 239). In conclusion, anti-hUBF antibodies were predominantly detected in patients with CTDs and UCTD. In patients with CTDs, SSc exhibited a high ratio, displaying a lower mRSS and higher incidence of ILD. In patients with UCTD, careful follow-up is recommended as they may develop CTDs in the future.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal , Autoanticorpos , Fatores de Transcrição , Humanos , Masculino , Feminino , Estudos Retrospectivos , Pessoa de Meia-Idade , Autoanticorpos/sangue , Autoanticorpos/imunologia , Adulto , Idoso , Doenças do Tecido Conjuntivo/imunologia , Doenças do Tecido Conjuntivo/diagnóstico , Escleroderma Sistêmico/imunologia , Escleroderma Sistêmico/sangue , Escleroderma Sistêmico/diagnóstico , Escleroderma Sistêmico/complicações , Índice de Gravidade de Doença , Doenças do Tecido Conjuntivo Indiferenciado/imunologia , Doenças do Tecido Conjuntivo Indiferenciado/complicaçõesRESUMO
To summarize the current therapies for skin cancers, the Japanese Skin Cancer Society issued the first guidelines for skin cancers, including melanoma, squamous cell carcinoma, basal cell carcinoma (BCC), and extramammary Paget's disease, in 2007. These guidelines were revised in 2015. Herein, we present the English version of the 2021 edition of the Japanese clinical guidelines for BCC. In the latest edition, all procedures were performed according to the Grading of Recommendations, Assessment, Development and Evaluation systems. The clinical questions that could not be answered were selected for further analysis. A comprehensive literature search, systematic review, and recommendations for each clinical question were determined by a multidisciplinary expert panel comprising dermatologists, a plastic and reconstructive surgeon, and a pathologist. Surgical resection is the gold-standard therapy of BCC. Radiotherapy or topical treatments, other than surgical resection, have been used in some cases. Patients with unresectable or metastatic BCC require systemic therapy. Novel agents, such as immune response modifiers or hedgehog pathway inhibitors, are emerging worldwide for the treatment of BCC. Based on these viewpoints, four relevant clinical questions regarding, surgical resection, radiotherapy, topical treatment, and systemic therapy, were raised in this report that aims to help clinicians select suitable therapies for their patients.
Assuntos
Carcinoma Basocelular , Neoplasias Cutâneas , Humanos , Carcinoma Basocelular/terapia , Carcinoma Basocelular/patologia , Carcinoma Basocelular/diagnóstico , Neoplasias Cutâneas/terapia , Neoplasias Cutâneas/patologia , Neoplasias Cutâneas/diagnóstico , Japão , Dermatologia/normas , Sociedades Médicas/normas , Antineoplásicos/uso terapêutico , População do Leste AsiáticoRESUMO
Mitochondrial F-ATP synthase produces the majority of ATP for cellular functions requiring free energy. The structural basis for proton motive force-driven rotational catalysis of ATP formation in the holoenzyme remains to be determined. Here, the purification and two-dimensional crystallization of bovine heart mitochondrial F-ATP synthase are reported. Two-dimensional crystals of up to 1 µm in size were grown by dialysis-mediated detergent removal from a mixture of decylmaltoside-solubilized 1,2-dimyristoyl-sn-glycero-3-phosphocholine and F-ATP synthase against a detergent-free buffer. A projection map calculated from an electron micrograph of a negatively stained two-dimensional crystal revealed unit-cell parameters of a = 185.0, b = 170.3 Å, γ = 92.5°.
Assuntos
Trifosfato de Adenosina/química , Mitocôndrias Cardíacas/química , ATPases Mitocondriais Próton-Translocadoras/química , Subunidades Proteicas/química , Animais , Bovinos , Cristalização , Cristalografia por Raios X , Dimiristoilfosfatidilcolina/análogos & derivados , Dimiristoilfosfatidilcolina/química , Microscopia Eletrônica , Mitocôndrias Cardíacas/enzimologia , ATPases Mitocondriais Próton-Translocadoras/isolamento & purificação , Conformação Proteica , Subunidades Proteicas/isolamento & purificaçãoRESUMO
Argonaute (AGO) proteins use small RNAs to recognize transcripts targeted for silencing in plants and animals. Many AGOs cleave target RNAs using an endoribonuclease activity termed 'slicing'. Slicing by DNA-guided prokaryotic AGOs has been studied in detail, but structural insights into RNA-guided slicing by eukaryotic AGOs are lacking. Here we present cryogenic electron microscopy structures of the Arabidopsis thaliana Argonaute10 (AtAgo10)-guide RNA complex with and without a target RNA representing a slicing substrate. The AtAgo10-guide-target complex adopts slicing-competent and slicing-incompetent conformations that are unlike known prokaryotic AGO structures. AtAgo10 slicing activity is licensed by docking target (t) nucleotides t9-t13 into a surface channel containing the AGO endoribonuclease active site. A ß-hairpin in the L1 domain secures the t9-t13 segment and coordinates t9-t13 docking with extended guide-target pairing. Results show that prokaryotic and eukaryotic AGOs use distinct mechanisms for achieving target slicing and provide insights into small interfering RNA potency.
Assuntos
Arabidopsis , Proteínas Argonautas , Animais , Proteínas Argonautas/metabolismo , RNA Interferente Pequeno/genética , Interferência de RNA , Arabidopsis/genética , Arabidopsis/metabolismo , Domínio Catalítico , RNA de Plantas/genética , RNA de Plantas/metabolismoRESUMO
BACKGROUND: Pain is a complex experience with both sensory and affective dimensions, and the affective dimension can increase the risks of chronic pain development. It is thus critical to identify factors influencing the affective dimension of pain. OBJECTIVE: This study aimed to identify the relationship between the affective dimension of pain and disorder site (primary pain source). METHODS: Study participants were recruited from patients referred for physical therapy at an orthopedic outpatient clinic. Pain quality including the affective dimension, disorder site from descriptive medical diagnosis, pain intensity, duration from pain onset, and demographic data, was collected. A multivariable logistic regression model was constructed to analyze the relationship between the affective dimension of pain and the disorder site. RESULTS: A total of 282 participants were included. Cervical and lumbar spine disorders were significantly associated with an affective dimension of pain compared to limbs disorders when adjusted for age, sex, pain intensity, and duration from the onset in the regression model. CONCLUSIONS: Regardless of duration from the onset and other confounding factors, disorder site is correlated with the affective dimension of pain. Multidimensional pain assessment is crucial when clinicians evaluate patients with cervical and lumbar spine disorders, even in the acute phase.
Assuntos
Dor Crônica , Humanos , Estudos Transversais , Medição da DorRESUMO
Skin wound healing is mediated by inflammatory cell infiltration of the wound site. Inducible costimulator (ICOS), expressed on activated T cells, and its ligand, ICOS ligand (ICOSL), expressed on antigen-presenting cells, have been considered a single receptor-ligand pair. Although the ICOS-ICOSL pathway participates in adaptive immunity, its roles in skin wound healing, which is mediated by innate immune responses, remain unknown. To clarify these roles, repair of excisional wounds was examined in ICOS(-/-) mice, ICOSL(-/-) mice, and ICOS(-/-)ICOSL(-/-) mice. Each mutant strain showed similar, dramatic delays in wound healing, especially at early times. Knockout mice showed suppressed keratinocyte migration, angiogenesis, and granulation tissue formation, and diminished T-cell, macrophage, and neutrophil infiltration. The loss of ICOS and/or ICOSL resulted in marked suppression of cytokine expression in wounds, especially the Th2 cytokines interleukin (IL)-4, IL-6, and IL-10. T-cell transfer experiments and T-cell depletion therapy further clarified the important roles of ICOS expressed on T cells and its interaction with ICOSL. Application of IL-6, but not IL-4, to the wounds significantly increased the onset of early wound healing in mutant mice. Thus, our results indicate that ICOS-ICOSL costimulatory signaling has critical roles during wound healing, most likely by inducing IL-6 production.
Assuntos
Citocinas/biossíntese , Imunidade Inata/imunologia , Proteína Coestimuladora de Linfócitos T Induzíveis/fisiologia , Transdução de Sinais/imunologia , Pele/imunologia , Cicatrização/imunologia , Adjuvantes Imunológicos/farmacologia , Animais , Células Apresentadoras de Antígenos/imunologia , Movimento Celular , Proliferação de Células , Citocinas/farmacologia , Tecido de Granulação/imunologia , Ligante Coestimulador de Linfócitos T Induzíveis/metabolismo , Interleucina-4/farmacologia , Interleucina-6/farmacologia , Queratinócitos/fisiologia , Ativação Linfocitária/imunologia , Transfusão de Linfócitos , Linfopenia/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Miofibroblastos/fisiologia , Neovascularização Fisiológica , Infiltração de Neutrófilos/imunologia , Pele/irrigação sanguínea , Linfócitos T/imunologiaRESUMO
The objectives of this study were to investigate the time-course of influence of compression of bone tunnel wall in ligament reconstruction on tissue around the bone tunnel and to histologically examine the mechanism of preventing the complication of bone tunnel dilation, using rabbit tibia. A model in which the femoral origin of the extensor digitorum longus tendon was cut and inserted into a bone tunnel made proximal to the tibia was prepared in the bilateral hind legs of 20 Japanese white rabbits. In each animal, a tunnel was made using a drill only in the right leg, while an undersized bone tunnel was made by drilling and then dilated by compression using a dilator to the same tunnel size as that in the right leg. Animals were sacrificed at 0, 2, 4, 8 and 12 weeks after surgery (4 animals at each time point). Observation of bone tunnels by X-ray radiography showed osteosclerosis in the 2- and 4-week dilation groups. Osteosclerosis appeared as white lines around the bone tunnel on X-ray radiography. This suggests that dilation promotes callus formation in the bone tunnel wall and prevents the complication of bone tunnel enlargement after ligament reconstruction.
RESUMO
Group IV materials are promising candidates for highly reliable and human-friendly thin-film thermoelectric generators, used for micro-energy harvesting. In this study, we investigated the synthesis and thermoelectric applications of a Ge-based ternary alloy thin film, Ge1-x-ySixSny. The solid-phase crystallization of the highly densified amorphous precursors allowed the formation of high-quality polycrystalline Ge1-x-ySixSny layers on an insulating substrate. The small compositions of Si and Sn in Ge1-x-ySixSny (x < 0.15 and y < 0.05) lowered the thermal conductivity (3.1 W m-1 K-1) owing to the alloy scattering of phonons, while maintaining a high carrier mobility (approximately 200 cm2 V-1 s-1). The solid-phase diffusion of Ga and P allowed us to control the carrier concentration to the order of 1019 cm-3 for holes and 1018 cm-3 for electrons. For both p- and n-type Ge1-x-ySixSny, the power factor peaked at x = 0.06 and y = 0.02, reaching 1160 µW m-1 K-2 for p-type and 2040 µW m-1 K-2 for n-type. The resulting dimensionless figure of merits (0.12 for p-type and 0.20 for n-type) are higher than those of most environmentally friendly thermoelectric thin films. These results indicate that group IV alloys are promising candidates for high-performance, reliable thin-film thermoelectric generators.
RESUMO
The human SMC5/6 complex is a conserved guardian of genome stability and an emerging component of antiviral responses. These disparate functions likely require distinct mechanisms of SMC5/6 regulation. In yeast, Smc5/6 is regulated by its Nse5/6 subunits, but such regulatory subunits for human SMC5/6 are poorly defined. Here, we identify a novel SMC5/6 subunit called SIMC1 that contains SUMO interacting motifs (SIMs) and an Nse5-like domain. We isolated SIMC1 from the proteomic environment of SMC5/6 within polyomavirus large T antigen (LT)-induced subnuclear compartments. SIMC1 uses its SIMs and Nse5-like domain to localize SMC5/6 to polyomavirus replication centers (PyVRCs) at SUMO-rich PML nuclear bodies. SIMC1's Nse5-like domain binds to the putative Nse6 orthologue SLF2 to form an anti-parallel helical dimer resembling the yeast Nse5/6 structure. SIMC1-SLF2 structure-based mutagenesis defines a conserved surface region containing the N-terminus of SIMC1's helical domain that regulates SMC5/6 localization to PyVRCs. Furthermore, SLF1, which recruits SMC5/6 to DNA lesions via its BRCT and ARD motifs, binds SLF2 analogously to SIMC1 and forms a separate Nse5/6-like complex. Thus, two Nse5/6-like complexes with distinct recruitment domains control human SMC5/6 localization.
Assuntos
Proteínas de Ciclo Celular , Proteínas Cromossômicas não Histona , Humanos , Proteínas de Ciclo Celular/metabolismo , Proteínas Cromossômicas não Histona/metabolismo , Proteômica , Compartimentos de Replicação ViralAssuntos
Anticorpos Monoclonais/efeitos adversos , Antineoplásicos/efeitos adversos , Melanoma/complicações , Pneumonia/diagnóstico , Pneumonia/etiologia , Idoso , Anticorpos Monoclonais/uso terapêutico , Antineoplásicos/uso terapêutico , Biomarcadores , Feminino , Humanos , Melanoma/tratamento farmacológico , Nivolumabe , Radiografia Torácica , Índice de Gravidade de Doença , Tomografia Computadorizada por Raios XRESUMO
BACKGROUND: Systemic sclerosis (SSc) is a systemic autoimmune disease characterized by excessive fibrosis. FcγRIIB is a low-affinity receptor for the Fc fragment of IgG. FcγRIIB is expressed on the surface of various leukocyte subsets and signals negative feedback pathways to down-regulate B-cell antigen receptor signaling. OBJECTIVE: The aim of the present study was to investigate the role of FcγRIIB in the development of a murine bleomycin-induced scleroderma model. METHODS: The experimental fibrosis model was generated by the intradermal injection of bleomycin into wild-type (WT) and FcγRIIB-deficient (FcγRIIB-/-) mice. We histologically assessed skin and lung fibrosis as well as inflammatory cell infiltration. Cytokine and chemokine expression levels were measured with RT-PCR. RESULTS: The severity of fibrosis in the skin and lung was significantly worse in FcγRIIB-/- mice than in WT mice. In the skin of bleomycin-treated mice, the numbers of CD8+ T cells, F4/80+ macrophages, MPO+ neutrophils, NK1.1+NK cells, and B220+ B cells were significantly higher in FcγRIIB-/- mice than in WT mice. The expression of TNF-α and IL-1ß was significantly higher in FcγRIIB-/- mice than in WT mice as was the expression of ICAM-1, CXCL2, and CCL3 in the affected skin. An adoptive transfer of splenic leukocytes from FcγRIIB-/- mice into WT mice showed exacerbated skin and lung fibrosis compared to WT mice without an adoptive transfer. CONCLUSION: These results indicate that FcγRIIB plays an inhibitory role in skin and lung fibrosis. Moreover, modulating FcγRIIB signaling has potential as a therapeutic approach for SSc.
Assuntos
Bleomicina , Escleroderma Sistêmico , Animais , Bleomicina/toxicidade , Linfócitos T CD8-Positivos/metabolismo , Modelos Animais de Doenças , Fibrose , Camundongos , Escleroderma Sistêmico/patologia , Pele/patologiaRESUMO
Sphingosine-1-phosphate (S1P) regulates numerous important physiological functions, including immune response and vascular integrity, via its cognate receptors (S1PR1 to S1PR5); however, it remains unclear how S1P activates S1PRs upon binding. Here, we determined the crystal structure of the active human S1PR3 in complex with its natural agonist S1P at 3.2-Å resolution. S1P exhibits an unbent conformation in the long tunnel, which penetrates through the receptor obliquely. Compared with the inactive S1PR1 structure, four residues surrounding the alkyl tail of S1P (the "quartet core") exhibit orchestrating rotamer changes that accommodate the moiety, thereby inducing an active conformation. In addition, we reveal that the quartet core determines G protein selectivity of S1PR3. These results offer insight into the structural basis of activation and biased signaling in G protein-coupled receptors and will help the design of biased ligands for optimized therapeutics.
RESUMO
Styrene maleic acid copolymers (SMA) form discoidal lipid nanoparticles (lipid nanodisks) that mimic plasma high-density lipoproteins. We have previously prepared and characterized lipid nanodisks composed of SMA and the neutral phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). In the present study, we tested whether the surface charges can alter the physicochemical and biological properties of lipid-SMA discoidal particles. Unlike the case of DMPC alone, addition of saline to the buffer was necessary to induce the formation of lipid-SMA complexes containing either 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG) or 1,2-dimyristoyl-3-trimethylammonium-propane (DMTAP), with formation efficiency being dependent on the concentration of charged lipids. After purification, DMPG- or DMTAP-containing discoidal particles with an approximate size of 10 nm were obtained in a manner similar to DMPC alone. Although DMPG and DMTAP appeared to be similarly incorporated into the lipid nanodisks, the zeta potentials of both particles were comparable. That is, no significant differences were observed in the physicochemical properties between the lipid-SMA nanodisks. Compared to DMPC-SMA nanodisks, the uptake of DMPG or DMTAP-containing discoidal particles by RAW264 cells was increased for both particle types, whereas in MDA-MB-231 cells, only DMTAP-containing discoidal particle uptake was increased. In addition, fluorescence microscopy revealed that lipid-SMA nanodisks are localized adjacent to the plasma membrane of RAW264 cells but in MDA-MB-231 cells they accumulated in the center of the cell. Furthermore, these particles caused cytotoxicity in a cell-type dependent manner, with high toxicity in MDA-MB-231. These results raised the possibility that compositional alterations in lipid-SMA discoidal particles may modulate biological reactions in vivo.