RESUMO
Tumor-associated macrophages are abundant infiltrating cells in the tumor microenvironment (TME). Macrophages can be classified into several types of subsets based on their immune responses. Among those subsets, M2 macrophages contribute to anti-inflammatory responses and create an immunosuppressive environment that promotes tumor cell proliferation. In a previous study, human cancer patients with high M2 macrophages showed a worse prognosis for many types of tumors. However, studies examining the relationship between M2 macrophages and clinical outcomes in canine tumors are limited. In the previous human and canine studies, CD204 has been used as the marker for detecting M2 macrophages. Then we evaluated CD204+ and total macrophages infiltration and its association with clinical outcomes in canine solid tumors. In this study, we examined dogs with oral malignant melanoma (OMM), pulmonary adenocarcinoma (PA), hepatocellular carcinoma (HCC), and transitional cell carcinoma (TCC). Compared to healthy tissues, CD204+ and total macrophages were increased in OMM, PA, and TCC, but not in HCC. High CD204+ macrophage levels were significantly associated with lung metastasis in TCC (P = 0.030). Kaplan-Meier analysis revealed that high CD204+ macrophage levels were associated with shorter overall survival (OS) in canine patients with PA (P = 0.012) and TCC (P = 0.0053). These results suggest that CD204+ macrophages contribute to tumor progression and could be a prognostic factor in dogs with PA and TCC.
Assuntos
Carcinoma Hepatocelular , Carcinoma de Células de Transição , Doenças do Cão , Neoplasias Hepáticas , Humanos , Cães , Animais , Macrófagos Associados a Tumor , Carcinoma Hepatocelular/veterinária , Carcinoma de Células de Transição/veterinária , Neoplasias Hepáticas/veterinária , Prognóstico , Microambiente TumoralRESUMO
This study investigated the long-term survival and incidence of secondary fractures after fragility hip fractures. The 5-year survival rate was 62%, and the mortality risk was seen in patients with GNRI < 92. The 5-year incidence of secondary fracture was 22%, which was significantly higher in patients with a BMI < 20. BACKGROUND: Malnutrition negatively influences the postoperative survival of patients with fragility hip fractures (FHFs); however, little is known about their association over the long term. OBJECTIVE: This study evaluated the ability of the geriatric nutritional risk index (GNRI) as a risk factor for long-term mortality after FHFs. METHODS: This study included 623 Japanese patients with FHFs over the age of 60 years. We prospectively collected data on admission and during hospitalization and assessed the patients' conditions after discharge through a questionnaire. We examined the long-term mortality and the incidence of secondary FHFs and assessed the prognostic factors. RESULTS: The mean observation period was 4.0 years (range 0-7 years). The average age at the time of admission was 82 years (range 60-101 years). The overall survival after FHFs (1 year, 91%; 5 years, 62%) and the incidence of secondary FHFs were high (1 year, 4%; 5 years, 22%). The multivariate Cox proportional hazard analysis revealed the risk factors for mortality as older age (hazard ratio [HR] 1.04), male sex (HR 1.96), lower GNRI score (HR 0.96), comorbidities (malignancy, HR 2.51; ischemic heart disease, HR 2.24; revised Hasegawa dementia scale ≤ 20, HR 1.64), no use of active vitamin D3 on admission (HR 0.46), and a lower Barthel index (BI) (on admission, HR 1.00; at discharge, HR 0.99). The GNRI scores were divided into four risk categories: major risk (GNRI, < 82), moderate risk (82-91), low risk (92-98), and no risk (> 98). Patients at major and moderate risks of GNRI had a significantly lower overall survival rate (p < 0.001). Lower body mass index (BMI) was also identified as a prognostic factor for secondary FHFs (HR 0.88 [p = 0.004]). CONCLUSIONS: We showed that older age, male sex, a lower GNRI score, comorbidities, and a lower BI are risk factors for mortality following FHFs. GNRI is a novel and simple predictor of long-term survival after FHFs.
Assuntos
Fraturas do Quadril , Desnutrição , Humanos , Masculino , Idoso , Pessoa de Meia-Idade , Idoso de 80 Anos ou mais , Avaliação Nutricional , Prognóstico , Desnutrição/complicações , Desnutrição/epidemiologia , Fraturas do Quadril/etiologia , Fatores de Risco , Avaliação Geriátrica , Estado Nutricional , Estudos RetrospectivosRESUMO
A 76-year-old Japanese man was admitted to hospital for treatment of fever and skin lesion at the implantation site of his pacemaker. During his hospitalization, vancomycin-intermediate Staphylococcus aureus (MIC 4 µg/mL) with reduced susceptibility to daptomycin was isolated from venous blood. This isolate was identified as methicillin-resistant S. aureus with SCCmec IV and was genotyped as sequence type 81, coa VIIa and spa type t7044, harbouring blaZ, aac(6')-aph(2â³) and enterotoxin(-like) genes sea, seb, sek, sel, selx and selw. The patient was successfully treated with daptomycin, minocycline and sulfamethoxazole/trimethoprim. We describe the identification of sequence type 81/SCCmec IV vancomycin-intermediate S. aureus from pacemaker-associated septicaemia.
RESUMO
Although antimicrobial peptides (AMPs) play an integral role in the regulation of intestinal microbiota and homeostasis, their expression in canine gastrointestinal diseases, including idiopathic inflammatory bowel disease (IBD) and intestinal lymphoma, remains unknown. The objective of this study was to investigate the intestinal expression of AMPs in dogs with IBD or intestinal lymphoma. IBD was diagnosed in 44 dogs, small cell intestinal lymphoma in 25 dogs, and large cell intestinal lymphoma in 19 dogs. Twenty healthy beagles were used as normal controls. Duodenal mRNA expression of six representative AMPs - lactoferrin, lysozyme, cathelicidin, secretory leukocyte peptidase inhibitor (SLPI), bactericidal/permeability increasing protein (BPI), and canine beta defensin (CBD103) - was quantified by real-time reverse transcription polymerase chain reaction. The relative expression of BPI, lactoferrin, and SLPI was significantly higher in dogs with IBD and intestinal lymphomas than in healthy controls. Interestingly, the expression patterns of AMPs differed between dogs with IBD and those with intestinal lymphomas, especially small cell lymphoma. Increased expression of BPI differentiated IBD from dogs with small cell intestinal lymphoma, with a sensitivity of 93.2%, a specificity of 100%, and an area under the curve of 0.955. These results suggest that the expression patterns of AMP aid in the diagnosis of canine IBD and intestinal lymphoma, although it remains uncertain whether the altered AMP expression is the cause or effect of mucosal inflammation.
Assuntos
Anti-Infecciosos/metabolismo , Peptídeos Catiônicos Antimicrobianos/genética , Doenças do Cão/genética , Duodeno/metabolismo , Doenças Inflamatórias Intestinais/veterinária , Neoplasias Intestinais/veterinária , Linfoma/veterinária , Animais , Peptídeos Catiônicos Antimicrobianos/biossíntese , Cães , Feminino , Expressão Gênica , Doenças Inflamatórias Intestinais/genética , Neoplasias Intestinais/genética , Linfoma/genética , MasculinoRESUMO
To elucidate if microglial P2Y12 receptor (P2Y12R) mechanisms are involved in the trigeminal spinal subnucleus caudalis (Vc; also known as the medullary dorsal horn) in intraoral cancer pain, we developed a rat model of tongue cancer pain. Squamous cell carcinoma (SCC) cells were inoculated into the tongue of rats; sham control rats received the vehicle instead. Nociceptive behavior was measured as the head-withdrawal reflex threshold (HWRT) to mechanical or heat stimulation applied to the tongue under light anesthesia. On day 14 after the SCC inoculation, activated microglia and P2Y12R expression were examined immunohistochemically in the Vc. The HWRT was also studied in SCC-inoculated rats with successive intra-cisterna magna (i.c.m.) administration of specific P2Y12R antagonist (MRS2395) or intraperitoneal administration of minocycline, a microglial activation inhibitor. Tongue cancer was histologically verified in SCC-inoculated rats, within which the HWRT to mechanical stimulation of the tongue was significantly decreased, as compared with that of vehicle-inoculated rats, although the HWRT to heat stimulation was not. Microglia was strongly activated on day 14, and the administration of MRS2395 or minocycline reversed associated nocifensive behavior and microglial activation in SCC-inoculated rats for 14 d. The activity of Vc wide dynamic range nociceptive neurons was also recorded electrophysiologically in SCC-inoculated and sham rats. Background activity and noxious mechanically evoked responses of wide dynamic range neurons were significantly increased in SCC-inoculated rats versus sham rats, and background activity and mechanically evoked responses were significantly suppressed following i.c.m. administration of MRS2395 in SCC-inoculated rats as compared with sham. The present findings suggest that SCC inoculation that produces tongue cancer results in strong activation of microglia via P2Y12 signaling in the Vc, in association with increased excitability of Vc nociceptive neurons, reflecting central sensitization and resulting in tongue mechanical allodynia.
Assuntos
Dor do Câncer/metabolismo , Carcinoma de Células Escamosas/metabolismo , Microglia/metabolismo , Neuralgia/metabolismo , Receptores Purinérgicos P2Y12/metabolismo , Neoplasias da Língua/metabolismo , Núcleo Espinal do Trigêmeo/metabolismo , Adenina/análogos & derivados , Adenina/farmacologia , Animais , Imuno-Histoquímica , Masculino , Minociclina/farmacologia , Nociceptores/metabolismo , Ratos , Ratos Endogâmicos F344 , Transdução de Sinais , Valeratos/farmacologiaRESUMO
Inferior alveolar nerve (IAN) injury induces persistent ectopic pain which spreads to a wide area in the orofacial region. Its exact mechanism remains unclear. We investigated the involvement of nitric oxide (NO) in relation to ectopic orofacial pain caused by IAN transection (IANX). We assessed the changes in mechanical sensitivity of the whisker pad skin following IANX, neuronal nitric oxide synthase (nNOS) expression in the trigeminal ganglion (TG), and the functional significance of NO in relation to the mechanical allodynia following intra-TG administration of a chemical precursor to NO and selective nNOS inhibitors. IANX induced mechanical allodynia, which was diminished by intra-TG administration of selective nNOS inhibitors. NO metabolites and nNOS immunoreactive neurons innervating the lower lip were also increased in the TG. Intra-TG administration of nNOS substrate induced the mechanical allodynia. The present findings suggest that NO released from TG neurons regulates the excitability of TG neurons innervating the whisker pad skin, and the enhancement of TG neuronal excitability may underlie ectopic mechanical allodynia.
Assuntos
Dor Facial/etiologia , Nervo Mandibular/fisiopatologia , Neuralgia/etiologia , Óxido Nítrico/fisiologia , Transdução de Sinais/fisiologia , Traumatismos do Nervo Trigêmeo/complicações , Animais , Arginina/análogos & derivados , Arginina/farmacologia , Inibidores Enzimáticos/farmacologia , Precursores Enzimáticos/farmacologia , Hiperalgesia/etiologia , Indazóis/farmacologia , Lábio/inervação , Masculino , Nervo Mandibular/patologia , Vias Neurais/patologia , Vias Neurais/fisiopatologia , Neurônios/efeitos dos fármacos , Neurônios/patologia , Óxido Nítrico/análise , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase Tipo I/análise , Óxido Nítrico Sintase Tipo I/antagonistas & inibidores , Limiar da Dor/fisiologia , Ratos , Ratos Sprague-Dawley , Limiar Sensorial/fisiologia , Transdução de Sinais/efeitos dos fármacos , Tato/fisiologia , Gânglio Trigeminal/efeitos dos fármacos , Gânglio Trigeminal/patologiaRESUMO
Angiogenesis plays a pivotal role in many pathological processes including chronic liver diseases. Various factors, such as renin-angiotensin-aldosterone system (RAAS), insulin resistance (IR), and reactive oxygen species (ROS) contribute reciprocally to promote angiogenesis. Blockade of RAAS by angiotensin-converting enzyme inhibitor (ACE-I) or angiotensin II (AngII) receptor blocker (ARB) markedly attenuates liver fibrosis and hepatocellular carcinoma (HCC) along with suppression of angiogenesis, IR, and ROS. Aldosterone (Ald), a downstream component of AngII, is also involved in these processes, and a selective Ald blocker (SAB) significantly suppressed the progression of chronic liver diseases. The IR status itself has shown to directly accelerate the progression of chronic liver diseases whereas inhibition of ROS by iron chelator suppressed it through augmentation and inhibition of neovascularization. The combination therapy of ACE-I/ARB/SAB with other clinically used agents, such as interferon, imatinib mesylate, vitamin K, iron chelator, and branched-chain amino acids (BCAA) exerted more potent inhibitory effects on the development of liver fibrosis and HCC than the treatment using a single agent alone. Collectively, the anti-angiogenic treatment targeting RAAS, IR, ROS with clinically available agents may become a new therapeutic strategy against the progression of chronic liver diseases.
Assuntos
Resistência à Insulina , Hepatopatias/fisiopatologia , Neovascularização Patológica/fisiopatologia , Estresse Oxidativo/fisiologia , Sistema Renina-Angiotensina/fisiologia , Antagonistas de Receptores de Angiotensina/uso terapêutico , Inibidores da Enzima Conversora de Angiotensina/uso terapêutico , Doença Crônica , Humanos , Fígado/irrigação sanguínea , Fígado/efeitos dos fármacos , Fígado/patologia , Hepatopatias/tratamento farmacológico , Hepatopatias/patologia , Modelos Biológicos , Neovascularização Patológica/prevenção & controle , Estresse Oxidativo/efeitos dos fármacos , Sistema Renina-Angiotensina/efeitos dos fármacosRESUMO
OBJECTIVE: To determine the association of distinct clinical subsets with myositis-specific autoantibodies (MSAs) towards anti-155/140-kDa polypeptides [anti-155/140 antibodies (Abs)], anti-140-kDa polypeptides (anti-140 Abs), and anti-aminoacyl tRNA synthetases (ARS Abs) in Japanese patients with dermatomyositis (DM). METHODS: We compared the clinical features and short-term prognoses of 30 DM patients whose serological status included these MSAs. The MSAs were determined by immunoprecipitation. RESULTS: Anti-155/140 Abs (n = 5), anti-140 Abs (n = 8), and anti-ARS Abs (n = 7) did not overlap each other. All of the anti-155/140 Ab-positive patients (n = 5) were complicated by malignancies, as were all of the anti-140 Ab-positive patients (n = 8), who showed rapidly progressive interstitial lung disease (ILD). The survival rate at 6 months from the diagnosis of DM was significantly lower in the anti-140 Ab-positive patients than in the other patients. CONCLUSION: This is the first study to report, in a single cohort of DM patients, that distinct clinical subsets are distributed in an anti-155/140 Ab-positive group, an anti-140 Ab-positive group, or an anti-ARS Ab-positive group. Our data also confirm previous evidence that anti-155/140 Abs are involved in malignancies and that anti-140 Abs are involved in rapidly progressive ILD.
Assuntos
Aminoacil-tRNA Sintetases/imunologia , Autoanticorpos/imunologia , Dermatomiosite/diagnóstico , Dermatomiosite/imunologia , RNA de Transferência/imunologia , Adulto , Idoso , Estudos de Coortes , Estudos Transversais , Feminino , Humanos , Imunoprecipitação , Japão , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Peptídeos/imunologia , Probabilidade , Estatísticas não ParamétricasRESUMO
BACKGROUND: Systemic sclerosis (SSc) is a connective tissue disorder with excessive fibrosis of the skin and various internal organs. Although SSc is a heterogeneous disease, it has been reported that the particular antinuclear antibodies (ANA) are often indicative of clinical features, disease course and overall severity. OBJECTIVE: To clarify the association of clinical and prognostic features with serum ANA in Japanese patients with SSc. METHODS: We studied 203 Japanese patients diagnosed with SSc, who visited our hospital during the period 1983-2005. Six SSc-related ANA were identified using indirect immunofluorescence, double immunodiffusion and immunoprecipitation assays. RESULTS: Patients with SSc were classified into six ANA-based subgroups and a group without ANA. As expected, antitopoisomerase I antibody (Ab, n=64), anti-RNA polymerases (RNAP) Ab (n=12) and anti-U3 RNP Ab (n=5) were associated with diffuse cutaneous SSc, whereas anticentromere Ab (ACA, n=75), anti-Th/To Ab (n=7) and anti-U1 RNP Ab (n=10) were frequently detected in patients with limited cutaneous SSc. Clinical features of the ANA-negative group (n=10) were heterogeneous. Consistent with previous findings in Caucasian and/or black African patients, antitopoisomerase I Ab was associated with the involvement of vascular and pulmonary fibrosis, leading to decreased survival rate. However, no patients with anti-RNAP Ab developed renal crisis and the frequency of isolated pulmonary hypertension in patients with ACA, anti-Th/To Ab or anti-U3 RNP Ab was similar to that in other ANA-based subgroups. CONCLUSION: These results indicate that the clinical relevance of SSc-related ANA in Japanese patients differs in some aspects from that in Caucasian and/or black African patients.
Assuntos
Anticorpos Antinucleares/genética , DNA Topoisomerases Tipo I/genética , Escleroderma Sistêmico/imunologia , Anticorpos Antinucleares/análise , Anticorpos Antinucleares/imunologia , Povo Asiático/etnologia , Causas de Morte , DNA Topoisomerases Tipo I/análise , Feminino , Fibrose , Humanos , Japão/etnologia , Masculino , Pessoa de Meia-Idade , Escleroderma Sistêmico/tratamento farmacológico , Taxa de SobrevidaRESUMO
OBJECTIVE: Myositis-specific autoantibodies (MSAs) are a useful tool in diagnosis, defining clinical subsets and predicting prognosis of dermatomyositis (DM) and polymyositis (PM). In this study, we identified a novel MSA reactive with 155 and 140 kDa nuclear proteins [anti-155/140 antibody (Ab)] and determined the clinical feature of DM patients positive for this autoantibody (autoAb). METHODS: Sera from 52 Japanese patients with DM, 9 with PM, 48 with systemic lupus erythematosus (SLE), 126 with systemic sclerosis and 18 with idiopathic interstitial pneumonia were examined by immunoprecipitation assays. Positive sera were further characterized by immunodepletion and immunofluorescence staining. RESULTS: Seven of the 52 (13%) Japanese patients with DM immunoprecipitated 155 and 140 kDa proteins from 35S-labelled K562 leukaemia cell extract. No patients with SLE, systemic sclerosis or idiopathic interstitial pneumonia as well as healthy controls were positive for this autoAb. Patients with anti-155/140 Ab developed heliotrope rash, Gottron's papules or sign and flagellate erythema significantly more frequently than those negative. Notably, internal malignancy was found at significantly higher frequency in those positive than those negative (71 vs 11%; P < 0.005). In contrast, none of these patients positive for this autoAb had interstitial lung disease. CONCLUSIONS: This novel MSA is associated with cancer-associated DM and may serve as a diagnostic serological marker for this specific subset.
Assuntos
Anticorpos Antinucleares/sangue , Dermatomiosite/diagnóstico , Neoplasias/complicações , Proteínas Nucleares/imunologia , Síndromes Paraneoplásicas/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Dermatomiosite/etiologia , Dermatomiosite/imunologia , Eletroforese em Gel de Poliacrilamida/métodos , Feminino , Humanos , Imunoprecipitação/métodos , Células K562 , Doenças Pulmonares Intersticiais/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Masculino , Pessoa de Meia-Idade , Neoplasias/imunologia , Síndromes Paraneoplásicas/etiologia , Síndromes Paraneoplásicas/imunologia , Escleroderma Sistêmico/imunologiaRESUMO
Ultraviolet (UV) irradiation induces damage of the skin, and in particular, photoageing is known to be the result of chronic UV irradiation. Many investigations have attempted to clarify the mechanisms of photoageing induced by chronic UVA irradiation, but consensus has not been achieved yet by in vivo experiments, mostly due to differences among UV sources and animals used for experiments. In vitro experiments have shown that a single exposure to UVA irradiation causes overexpression of matrix metalloproteinases and denaturation of collagen, but the mechanisms of the photoageing effects of chronic UVA irradiation are still unclear. To examine the effects of chronic UVA irradiation, we used an in vitro fibroblast cellular ageing system as a model of photoageing. Chronic UVA irradiation of normal human fibroblasts induced shortening of the cellular life span and an increase of cellular diameter, in parallel with expression of senescence-associated beta-galactosidase. Extracellular degradation enzyme, matrix metalloproteinase 1 (MMP-1) was overexpressed after repeated UVA irradiation, but tissue inhibitor of metalloproteinase 1 (TIMP-1) expression was hardly changed by chronic UVA irradiation. We conclude that chronic UVA irradiation of normal human fibroblasts induces cellular functional changes, leading to accelerated cellular ageing and MMP-1 overexpression.
Assuntos
Senescência Celular/efeitos da radiação , Derme/efeitos da radiação , Envelhecimento da Pele/efeitos da radiação , Biomarcadores/análise , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Células Cultivadas , Derme/citologia , Derme/enzimologia , Fibroblastos/citologia , Fibroblastos/enzimologia , Fibroblastos/efeitos da radiação , Humanos , Metaloproteinase 1 da Matriz/genética , RNA Mensageiro/análise , Espécies Reativas de Oxigênio/farmacologia , Coloração e Rotulagem , Inibidor Tecidual de Metaloproteinase-1/genética , beta-Galactosidase/análiseRESUMO
STUDY DESIGN: Case report. OBJECTIVE: To report a rare thoracic intervertebral disc herniation followed by acutely progressing paraplegia. SETTING: Spinal Injuries Center, Fukuoka, Japan. METHOD: A 37-year-old man presented with sudden severe backache and acutely progressing motor impairments of both lower extremities after antecedent backache lasting about 5 days. Neurological examination showed analgesia and hypoesthesia below the T4 dermatome level, dysesthesia to pinprick below right inguinal level, and severe motor impairments of the lower extremities (Frankel classification C). Magnetic resonance (MR) imaging demonstrated spinal cord compression due to a postero-laterally existing epidural mass at the T2-T3 level. After laminectomy at the T2-T3 level, the sequestrated disc material was detected and excised as one piece through the right side of the dura. The excised herniated mass had a ring-like form and was thought to originate from the annulus fibrosis. RESULT: After the emergency surgery, he had complete relief from the backache and control of both lower extremities recovered gradually. At 4 weeks after the emergent operation, motor power of both lower extremities recovered almost completely. He was able to walk without any assistance. MR imaging study after surgery did not reveal the sequestrated mass, except for a mild disc bulging at the T2-T3 level. CONCLUSION: Accurate diagnosis of acute symptomatic thoracic disc herniation is occasionally difficult. However, timely and successful surgery could result in complete symptom relief and satisfactory results.
Assuntos
Deslocamento do Disco Intervertebral/diagnóstico , Deslocamento do Disco Intervertebral/etiologia , Paraplegia/complicações , Paraplegia/diagnóstico , Doença Aguda , Adulto , Progressão da Doença , Humanos , Deslocamento do Disco Intervertebral/cirurgia , Imageamento por Ressonância Magnética , Masculino , Paraplegia/cirurgia , Vértebras Torácicas , Resultado do TratamentoRESUMO
In the present study, we investigated the effects of levomepromazine on plasma risperidone concentrations in a steady state. Twenty patients taking risperidone at a stable dose for more than 2 weeks who were considered to require levomepromazine coadministration were selected. The scores of excitement in BPRS significantly decreased 2 weeks after the coadministration of levomepromazine. Plasma risperidone concentrations and the ratio of risperidone and 9-hydroxyrisperidone (risperidone/9-hydroxyrisperidone) did not change between before and 2 weeks after the coadministration of levomepromazine. The extrapyramidal symptoms were not worsened by the coadministration of levomepromazine. These results suggest that a low dosage of levomepromazine, use as a sedative adjuvant to risperidone treatment, have no statistically significant effect on the trough plasma concentrations of risperidone.
Assuntos
Antipsicóticos/sangue , Metotrimeprazina/farmacologia , Metotrimeprazina/uso terapêutico , Transtornos Psicóticos/tratamento farmacológico , Risperidona/sangue , Adolescente , Adulto , Antipsicóticos/metabolismo , Escalas de Graduação Psiquiátrica Breve , Relação Dose-Resposta a Droga , Feminino , Humanos , Masculino , Metotrimeprazina/administração & dosagem , Pessoa de Meia-Idade , Risperidona/metabolismoRESUMO
Effects of rice bran agglutinin (RBA) on human monoblastic leukemia U937 cells were examined in comparison with those of wheat germ agglutinin (WGA) and Viscum album agglutinin (VAA). These lectins inhibit cell growth, and several lines of evidence indicate that the growth inhibition is caused by the induction of apoptosis. We observed that RBA induces chromatin condensation, externalization of membrane phosphatidylserine, and DNA ladder formation, features of apoptosis. DNA ladder formation was inhibited by a general inhibitor against caspases, which are known to play essential roles in apoptosis. Flow cytometric analysis revealed that RBA and WGA cause G2/M phase cell cycle arrest with increased expression of Waf1/p21, while cell cycle arrest was not observed for VAA. These data indicate that RBA induces apoptosis associated with cell cycle arrest in U937 cells, and suggest that the induction mechanism for RBA is similar to that for WGA, but different from that for VAA.
Assuntos
Apoptose , Cromatina/efeitos dos fármacos , Ciclinas/agonistas , Fragmentação do DNA/efeitos dos fármacos , Lectinas/farmacologia , Lectinas de Plantas , Preparações de Plantas , Proteínas de Plantas/farmacologia , Ciclo Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Inibidor de Quinase Dependente de Ciclina p21 , Ciclinas/biossíntese , Inibidores de Cisteína Proteinase/farmacologia , Fase G2/efeitos dos fármacos , Humanos , Linfoma Difuso de Grandes Células B , Mitose/efeitos dos fármacos , Fosfatidilserinas/metabolismo , Proteínas Inativadoras de Ribossomos Tipo 2 , Toxinas Biológicas/farmacologia , Células U937 , Aglutininas do Germe de Trigo/farmacologiaRESUMO
We describe a patient in whom pemphigus foliaceus developed after cutaneous squamous cell carcinoma (SCC) metastasized to regional lymph nodes. Immunologic analysis revealed that production of anti-desmoglein 1 autoantibodies started when SCC metastasized, and the SCC expressed desmoglein 1, suggesting a pathogenic role of metastasized SCC in developing pemphigus foliaceus.
Assuntos
Caderinas/análise , Carcinoma de Células Escamosas/secundário , Metástase Linfática , Pênfigo/etiologia , Neoplasias Cutâneas/patologia , Idoso , Autoanticorpos/análise , Caderinas/biossíntese , Desmogleína 1 , Feminino , Humanos , Pênfigo/patologiaRESUMO
Although tumor necrosis factor receptor-associated factor 6 (TRAF6) is required in receptor activator of NF-kappaB-receptor activator of NF-kappaB ligand (RANK-RANKL) signaling for osteoclastogenesis, it has remained unclear whether TRAF6 is crucial in tumor necrosis factor alpha (TNF-alpha)-induced osteoclastogenesis. We examined TRAF6 function in the TNF-alpha-induced osteoclastogenesis by using osteoclast progenitor cells from TRAF6-deficient mice. The results indicated that TNF-alpha did not effectively induce osteoclast differentiation from osteoclast progenitor cells derived from these mice into mature multinucleated osteoclasts, although c-jun N-terminal kinase (JNK) and TNF-alpha activation was observed in osteoclast progenitor cells. Thus, we have provided the first evidence showing that TRAF6 is involved in TNF-alpha-induced osteoclastogenesis.
Assuntos
Osteoblastos/citologia , Osteogênese/fisiologia , Proteínas/metabolismo , Receptores do Fator de Necrose Tumoral/metabolismo , Transdução de Sinais , Células-Tronco/citologia , Fator de Necrose Tumoral alfa/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Proteínas Quinases JNK Ativadas por Mitógeno , Camundongos , Camundongos Knockout , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Proteínas/genética , Proteínas/fisiologia , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismo , Fator 6 Associado a Receptor de TNF , Fator de Necrose Tumoral alfa/farmacologia , Regulação para CimaRESUMO
The antimutagenic activities of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were examined by studying their effects on induction of 6-thioguanine (6TG)-resistant mutations by ethyl methanesulfonate (EMS) in cultured Chinese hamster V79 cells. DRA had a remarkable inhibitory effect against the cytotoxicity of EMS, when cells were simultaneously-treated with EMS, showing a blocking or scavenging activity of DHA in reduction of surviving fraction of cells. DHA had not so significant effect, when cells were treated before and after treatment with EMS. On the other hand, EPA had marked inhibiting effects against cytotoxicity of EMS, when cells were treated with EPA, before, simultaneous and after treatment with EMS. Against the induction of mutations by EMS, an antimutagenic activity of DHA was found when cells were pre-treated, simultaneously-treated or post-treated with DHA. EPA was also effective in reducing EMS-induced 6TG-resistant mutations when the cells were treated using the three different treatment procedures described above. The results suggest that in cultured Chinese hamster V79 cells, DHA and EPA may have both desmutagenic activity, which inactivates EMS chemically and/or enzymatically and bio-antimutagenic activity which suppresses mutation fixation after DNA is damaged by EMS.
Assuntos
Antimutagênicos/farmacologia , Ácidos Docosa-Hexaenoicos/farmacologia , Ácido Eicosapentaenoico/farmacologia , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Ensaio de Unidades Formadoras de Colônias , Cricetinae , Metanossulfonato de Etila/antagonistas & inibidores , Metanossulfonato de Etila/toxicidade , Testes de Mutagenicidade , Tioguanina/farmacologiaRESUMO
In order to investigate the mechanism of the production of oxidative DNA damage by hyperglycemia, we measured formamidopyrimidine N-glycosylase (FPG)-sensitive sites by the comet assay in human umbilical vein endothelial cells (HUVECs) cultured under various conditions including high glucose. Mean values of FPG-sensitive sites were higher in HUVECs cultured for 5 days in high glucose (45 mM) compared with normal glucose (5mM) medium (P<0.001). FPG-sensitive sites increased in a time-dependent manner under high glucose treatment (3 days: P<0.05, 5 days: P<0.001), whereas L-glucose, which is taken up poorly into the cells, gave a slight increase in FPG-sensitive sites (P<0.05). Flow cytometric analysis using 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate, di(acetoxymethyl ester) showed that incubation with L-glucose produced more reactive oxygen species than incubation with D-glucose. However, these increases were slight (1.22- and 1.12-folds, respectively). Incubation of HUVECs with aminoguanidine (100 microM) or pyridoxamine (1mM), which are inhibitors of glycation, decreased the levels of FPG-sensitive sites (P<0.001). However, these inhibitors did not suppress the intracellular generation of reactive oxygen species induced by high glucose. These results indicate that FPG-sensitive sites induced by high glucose are not due to intracellular reactive oxygen species. In order to clarify what caused the induction of FPG-sensitive sites, we investigated the effect of glyoxal and 3-deoxyglucosone (3-DG) on the induction of FPG-sensitive sites and the intracellular production of reactive oxygen species in HUVECs. Glyoxal and 3-DG at a concentration of 100 microg/ml induced FPG-sensitive sites (P<0.001, P<0.01, respectively). In contrast, glyoxal did not generate reactive oxygen species inside HUVECs. The results shown in this study suggest that glyoxal formed intracellularly or extracellularly during high glucose treatment might induce FPG-sensitive sites by a mechanism not involving reactive oxygen species.
Assuntos
Dano ao DNA , DNA/metabolismo , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , Glucose/toxicidade , Sítios de Ligação/efeitos dos fármacos , Células Cultivadas , Ensaio Cometa , DNA-Formamidopirimidina Glicosilase , Relação Dose-Resposta a Droga , Endotélio Vascular/citologia , Citometria de Fluxo , Guanidinas/farmacologia , Humanos , N-Glicosil Hidrolases/metabolismo , Oxirredução/efeitos dos fármacos , Piridoxamina/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Veias UmbilicaisRESUMO
Epigallocatechin gallate (EGCG) is known to induce apoptosis in various types of tumor cells, but the precise mechanism by which EGCG induces apoptosis remains to be elucidated. The Fas-Fas ligand system is one of the major pathways operating in the apoptotic cascade. The aim of this study was to examine the possibility that EGCG-binding to Fas triggers the Fas-mediated apoptosis. The EGCG treatment of human monocytic leukemia U937 cells resulted in elevation of caspase 8 activity and fragmentation of caspase 8. The DNA ladder formation caused by the EGCG treatment was inhibited by the caspase 8 inhibitor. These findings suggested the involvement of the Fas-mediated cascade in the EGCG-induced apoptosis in U937 cells. Affinity chromatography revealed the binding between EGCG and Fas. Thus, the results suggest that EGCG-binding to Fas, presumably on the cell surface, triggers the Fas-mediated apoptosis in U937 cells.
Assuntos
Apoptose , Catequina/metabolismo , Receptor fas/metabolismo , Apoptose/efeitos dos fármacos , Western Blotting , Caspase 8 , Caspase 9 , Inibidores de Caspase , Caspases/metabolismo , Catequina/análogos & derivados , Catequina/farmacologia , Cromatografia de Afinidade , Inibidores de Cisteína Proteinase/farmacologia , Relação Dose-Resposta a Droga , Ativação Enzimática/efeitos dos fármacos , Citometria de Fluxo , Humanos , Ligação Proteica/efeitos dos fármacos , Ligação Proteica/fisiologia , Células U937RESUMO
Resveratrol and quercetin are polyphenolic compounds found in grapes, red wine and other food products. In this study, we examined the effect of resveratrol and quercetin on the inhibition of angiogenesis in vitro. Resveratrol and quercetin inhibited the growth of bovine aorta endothelial (BAE) cells in a concentration-dependent manner (6-100 microM).The migration of BAE was obviously inhibited by resveratrol and weakly inhibited by quercetin. When the lengths of all tubes constructed in the 3-D culture system with or without resveratrol or quercetin were measured, resveratrol was found to inhibit the tube formation of BAE cells in a concentration-dependent manner (6-100 microM). On the other hand, quercetin at concentrations above 100 microM significantly inhibited the tube formation of vascular endothelial cells. From these results, we suggest that resveratrol and quercetin may prove useful in the development of useful therapeutic agents or preventive food factors for tumor angiogenesis.