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BACKGROUND: TLR7 is a key player in the antiviral immunity. TLR7 signaling activates antigen-presenting cells including DCs and macrophages. This activation results in the adaptive immunity including T cells and B cells. Therefore, TLR7 is an important molecule of the immune system. Based on these observations, TLR7 agonists considered to become a therapy weaponize the immune system against cancer. Radiation therapy (RT) is one of the standard cancer therapies and is reported to modulate the tumor immune response. In this study, we aimed to investigate the anti-tumor activity in combination of TLR7 agonist, DSP-0509, with RT and underlying mechanism. RESULT: We showed that anti-tumor activity is enhanced by combining RT with the TLR7 agonist DSP-0509 in the CT26, LM8, and 4T1 inoculated mice models. We found that once- weekly (q1w) dosing of DSP-0509 rather than biweekly (q2w) dosing is needed to achieve superior anti-tumor activities in CT26 model. Spleen cells from the mice in RT/DSP-0509 combination treatment group showed increased tumor lytic activity, inversely correlated with tumor volume, as measured by the chromium-release cytotoxicity assay. We also found the level of cytotoxic T lymphocytes (CTLs) increased in the spleens of completely cured mice. When the mice completely cured by combination therapy were re-challenged with CT26 cells, all mice rejected CT26 cells but accepted Renca cells. This rejection was not observed with CD8 depletion. Furthermore, levels of splenic effector memory CD8 T cells were increased in the combination therapy group. To explore the factors responsible for complete cure by combination therapy, we analyzed peripheral blood leukocytes (PBLs) mRNA from completely cured mice. We found that Havcr2low, Cd274low, Cd80high, and Il6low were a predictive signature for the complete response to combination therapy. An analysis of tumor-derived mRNA showed that combination of RT and DSP-0509 strongly increased the expression of anti-tumor effector molecules including Gzmb and Il12. CONCLUSION: These data suggest that TLR7 agonist, DSP-0509, can be a promising concomitant when used in combination with RT by upregulating CTLs activity and gene expression of effector molecules. This combination can be an expecting new radio-immunotherapeutic strategy in clinical trials.
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Receptor Toll-Like 7 , Animales , Receptor Toll-Like 7/agonistas , Ratones , Línea Celular Tumoral , Femenino , Activación de Linfocitos/efectos de los fármacos , Ratones Endogámicos BALB C , Glicoproteínas de Membrana/agonistas , Terapia Combinada , Humanos , Ratones Endogámicos C57BL , Neoplasias/terapia , Neoplasias/inmunología , Neoplasias/tratamiento farmacológico , Modelos Animales de Enfermedad , Linfocitos T/inmunología , Linfocitos T/efectos de los fármacosRESUMEN
Japan is lagging in cervical cancer prevention. The effectiveness of a self-sampling human papillomavirus (HPV) test, a possible measure to overcome this situation, has not yet been evaluated. A randomized controlled trial was performed to evaluate the effectiveness of a self-sampling HPV test on detection of cervical intraepithelial neoplasia grade 2 or worse (CIN2+) and screening uptake. Women between 30 and 58 years old who did not participate in the cervical cancer screening program for ≥3 years were eligible and assigned to the intervention group (cytology or self-sampling HPV test) or control group (cytology). Participants assigned to the intervention group were sent a self-sampling kit according to their ordering (opt-in strategy). A total of 7337 and 7772 women were assigned to the intervention and control groups, respectively. Screening uptake in the intervention group was significantly higher than that in the control group (20.0% vs. 6.4%; risk ratio: 3.10; 95% confidence interval [CI]: 2.82, 3.42). The compliance rate with cytology triage for HPV-positive women was 46.8% (95% CI: 35.5%, 58.4%). CIN2+ was detected in five and four participants in the intervention and control groups, respectively; there was no difference for intention-to-screen analysis (risk ratio: 1.32; 95% CI: 0.36, 4.93). Self-sampling of HPV test increased screening uptake; however, no difference was observed in the detection of CIN2+, probably due to the low compliance rate for cytology triage in HPV-positive women. Efforts to increase cytology triage are essential to maximize precancer detections.
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Detección Precoz del Cáncer , Infecciones por Papillomavirus , Displasia del Cuello del Útero , Neoplasias del Cuello Uterino , Humanos , Femenino , Persona de Mediana Edad , Adulto , Infecciones por Papillomavirus/diagnóstico , Infecciones por Papillomavirus/virología , Infecciones por Papillomavirus/epidemiología , Japón/epidemiología , Neoplasias del Cuello Uterino/diagnóstico , Neoplasias del Cuello Uterino/virología , Detección Precoz del Cáncer/métodos , Displasia del Cuello del Útero/diagnóstico , Displasia del Cuello del Útero/virología , Displasia del Cuello del Útero/epidemiología , Papillomaviridae/aislamiento & purificación , Frotis Vaginal/métodos , Manejo de Especímenes/métodos , Tamizaje Masivo/métodos , Lesiones Precancerosas/diagnóstico , Lesiones Precancerosas/virología , Virus del Papiloma HumanoRESUMEN
PURPOSE: In terms of medical policy for cervical cancer prevention, Japan lags far behind other industrialized countries. We initiated a randomized controlled trial to evaluate the self-sampling human papillomavirus (HPV) test as a tool to raise screening uptake and detection of pre-cancer. This study was conducted to explore the acceptability and preference of self-sampling using a subset of the data from this trial. METHODS: A pre-invitation letter was sent to eligible women, aged 30-59 years who had not undergone cervical cancer screening for three or more years. After excluding those who declined to participate in this trial, the remaining women were assigned to the self-sampling and control groups. A second invitation letter was sent to the former group, and those wanting to undergo the self-sampling test ordered the kit. A self-sampling HPV kit, consent form, and a self-administered questionnaire were sent to participants who ordered the test. RESULTS: Of the 7,340 participants in the self-sampling group, 1,196 (16.3%) administered the test, and 1,192 (99.7%) answered the questionnaire. Acceptability of the test was favorable; 75.3-81.3% of participants agreed with positive impressions (easy, convenient, and clarity of instruction), and 65.1-77.8% disagreed with negative impressions (painful, uncomfortable, and embarrassing). However, only 21.2% were confident in their sampling procedure. Willingness to undergo screening with a self-collected sample was significantly higher than that with a doctor-collected sample (89.3% vs. 49.1%; p<0.001). Willingness to undergo screening with a doctor-collected sample was inversely associated with age and duration without screening (both p<0.001), but that with a self-collected sample was not associated. CONCLUSIONS: Among women who used the self-sampling HPV test, high acceptability was confirmed, while concerns about self-sampling procedures remained. Screening with a self-collected sample was preferred over a doctor-collected sample and the former might alleviate disparities in screening rates.
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Infecciones por Papillomavirus , Neoplasias del Cuello Uterino , Femenino , Humanos , Frotis Vaginal/métodos , Virus del Papiloma Humano , Detección Precoz del Cáncer/métodos , Infecciones por Papillomavirus/diagnóstico , Japón , Papillomaviridae , Autocuidado/métodos , Manejo de Especímenes/métodos , Encuestas y Cuestionarios , Tamizaje Masivo/métodosRESUMEN
TLR7 is an innate immune receptor that recognizes single-stranded RNAs, and its activation leads to anti-tumor immune effects. Although it is the only approved TLR7 agonist in cancer therapy, imiquimod is allowed to be administered with topical formulation. Thus, systemic administrative TLR7 agonist is expected in terms of expanding applicable cancer types. Here, we demonstrated the identification and characterization of DSP-0509 as a novel small-molecule TLR7 agonist. DSP-0509 is designed to have unique physicochemical features that could be administered systemically with a short half-life. DSP-0509 activated bone marrow-derived dendritic cells (BMDCs) and induced inflammatory cytokines including type I interferons. In the LM8 tumor-bearing mouse model, DSP-0509 reduced tumor growth not only in subcutaneous primary lesions but also in lung metastatic lesions. DSP-0509 inhibited tumor growth in several syngeneic tumor-bearing mouse models. We found that the CD8+ T cell infiltration of tumor before treatment tended to be positively correlated with anti-tumor efficacy in several mouse tumor models. The combination of DSP-0509 with anti-PD-1 antibody significantly enhanced the tumor growth inhibition compared to each monotherapy in CT26 model mice. In addition, the effector memory T cells were expanded in both the peripheral blood and tumor, and rejection of tumor re-challenge occurred in the combination group. Moreover, synergistic anti-tumor efficacy and effector memory T cell upregulation were also observed for the combination with anti-CTLA-4 antibody. The analysis of the tumor-immune microenvironment by using the nCounter assay revealed that the combination of DSP-0509 with anti-PD-1 antibody enhanced infiltration by multiple immune cells including cytotoxic T cells. In addition, the T cell function pathway and antigen presentation pathway were activated in the combination group. We confirmed that DSP-0509 enhanced the anti-tumor immune effects of anti-PD-1 antibody by inducing type I interferons via activation of dendritic cells and even CTLs. In conclusion, we expect that DSP-0509, a new TLR7 agonist that synergistically induces anti-tumor effector memory T cells with immune checkpoint blockers (ICBs) and can be administered systemically, will be used in the treatment of multiple cancers.
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Inhibidores de Puntos de Control Inmunológico , Interferón Tipo I , Neoplasias , Receptor Toll-Like 7 , Animales , Ratones , Adyuvantes Inmunológicos/farmacología , Modelos Animales de Enfermedad , Inhibidores de Puntos de Control Inmunológico/farmacología , Inhibidores de Puntos de Control Inmunológico/uso terapéutico , Neoplasias/tratamiento farmacológico , Receptor Toll-Like 7/agonistas , Microambiente TumoralRESUMEN
A self-sampling human papillomavirus (HPV) test could improve the morbidity and mortality of cervical cancer in Japan. However, its effectiveness and feasibility have not been demonstrated sufficiently. Hence, we launched a randomized controlled trial, which is ongoing, and report the results of a secondary analysis. To ensure autonomous participation with a minimum selection bias, opt-out consent was obtained from women who met the inclusion criteria, and written consent was obtained from those who underwent a self-sampling test. The number of women who met the inclusion criteria was 20,555; 4283 and 1138 opted out before and after the assignment, respectively. Of the 7340 women in the self-sampling arm, 1372 (18.7%) ordered and 1196 (16.3%) underwent the test. Younger women in their 30 s and 40 s tended to undertake the test more frequently than older women in their 50 s (P for trend < 0.001). Invalid HPV test results were rare (1.3%), and neither adverse events nor serious complaints were reported. Despite adopting the opt-out procedure, more women than expected declined to participate, suggesting the need for a waiver of consent or assignment before consent to reduce selection bias. A self-sampling HPV test can be implemented in Japan and would be more accessible to young women, the predominant group affected by cervical cancer.
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Infecciones por Papillomavirus , Neoplasias del Cuello Uterino , Anciano , Detección Precoz del Cáncer/métodos , Femenino , Humanos , Japón , Tamizaje Masivo/métodos , Papillomaviridae , Autocuidado , Manejo de Especímenes , Frotis Vaginal/métodosRESUMEN
INTRODUCTION: Recently, the incidence of cervical cancer has increased in Japan, probably because of an interruption in human papillomavirus (HPV) vaccination and a low cervical cancer screening rate. There is a lack of evidence for self-sampling HPV testing as a cervical cancer screening tool in Japan. The Accelerating Cervical Cancer Elimination by Self-Sampling test trial aims to compare the effectiveness of screening using the self-sampling HPV test with that of routine screening concerning screening uptake and precancer detection. METHODS AND ANALYSIS: This trial has a single-municipality, open-label, parallel, superiority and randomised design. Approximately 20 000 women who have not undergone cervical cancer screening for at least 3 years will be assigned randomly to the self-sampling arm and the control arm using a 1:1 ratio. Participants assigned to the control arm will undergo routine cervical cancer screening (cytology test) provided by Ichihara City, while those assigned to the self-sampling arm will choose the routine screening or self-sampling HPV test. HPV tests will be performed using the cobas 8800 system (Roche Diagnostics, Rotkreuz, Switzerland). Participants who will undergo the self-sampling HPV testing will be recommended to undergo routine screening. The results of the cytology test and further tests, such as colposcopy and biopsy, will be collected and used for this trial. The risk ratio and risk difference in the proportion of participants with cervical intraepithelial neoplasia two or worse between the two arms will be calculated. The test for the null hypothesis (the detection rates are equal between the two arms) will be performed using Pearson's χ2 test. ETHICS AND DISSEMINATION: This trial was approved by the Research Ethics Committees of the Chiba Foundation for Health Promotion and Disease Prevention and the collaborating research institutes. The results will be disseminated through peer-reviewed journals and conference presentations. TRIAL REGISTRATION NUMBER: jRCT1030200276. Pre-results.
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Alphapapillomavirus , Infecciones por Papillomavirus , Neoplasias del Cuello Uterino , Detección Precoz del Cáncer/métodos , Femenino , Humanos , Tamizaje Masivo/métodos , Papillomaviridae , Infecciones por Papillomavirus/epidemiología , Ensayos Clínicos Controlados Aleatorios como Asunto , Neoplasias del Cuello Uterino/diagnóstico , Neoplasias del Cuello Uterino/patología , Neoplasias del Cuello Uterino/prevención & control , Frotis VaginalRESUMEN
Strategies to augment anti-cancer immune responses have recently demonstrated therapeutic utility. To date clinical success has been achieved through targeting co-inhibitory checkpoints such as CTLA-4, PD-1, and PD-L1. However, approaches that target co-activatory pathways are also being actively being developed. Here we report that the novel TLR7-selective agonist DSR-29133 is well tolerated in mice and leads to acute immune activation. Administration of DSR-29133 leads to the induction of IFNα/γ, IP-10, TNFα, IL-1Ra and IL-12p70, and to a reduction in tumor burden in syngeneic models of renal cancer (Renca), metastatic osteosarcoma (LM8) and colorectal cancer (CT26). Moreover, we show that the efficacy of DSR-29133 was significantly improved when administered in combination with low-dose fractionated radiotherapy (RT). Effective combination therapy required weekly administration of DSR-29133 commencing on day 1 of a fractionated RT treatment cycle, whereas no enhancement of radiation response was observed when DSR-29133 was administered at the end of the fractionated RT cycle. Combined therapy resulted in curative responses in a high proportion of mice bearing established CT26 tumors which was dependent on the activity of CD8+ T-cells but independent of CD4+ T-cells and NK/NKT cells. Moreover, long-term surviving mice originally treated with DSR-29133 and RT were protected by a tumor-specific memory immune response which could prevent tumor growth upon rechallenge. These results demonstrate that DSR-29133 is a potent selective TLR7 agonist that when administered intravenously can induce anti-tumor immune responses that can be further enhanced through combination with low-dose fractionated RT.
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Adenina/análogos & derivados , Antineoplásicos/farmacología , Quimioradioterapia/métodos , Neoplasias Experimentales/tratamiento farmacológico , Receptor Toll-Like 7/agonistas , Adenina/farmacología , Administración Intravenosa , Animales , Linfocitos T CD8-positivos/efectos de los fármacos , Linfocitos T CD8-positivos/inmunología , Fraccionamiento de la Dosis de Radiación , Humanos , Activación de Linfocitos/efectos de los fármacos , Ratones , Neoplasias Experimentales/inmunología , Neoplasias Experimentales/radioterapiaRESUMEN
Systemic administration of small molecule toll-like receptor (TLR)-7 agonists leads to potent activation of innate immunity and to the generation of anti-tumor immune responses. However, activation of TLRs with small molecule agonists may lead to the induction of TLR tolerance, defined as a state of hyporesponsiveness to subsequent agonism, which may limit immune activation, the generation of anti-tumor responses and clinical response. Our data reveal that dose scheduling impacts on the efficacy of systemic therapy with the selective TLR7 agonist, 6-amino-2-(butylamino)-9-((6-(2-(dimethylamino)ethoxy)pyridin-3-yl)methyl)-7,9-dihydro-8H-purin-8-one (DSR-6434). In a preclinical model of renal cell cancer, systemic administration of DSR-6434 dosed once weekly resulted in a significant anti-tumor response. However, twice weekly dosing of DSR-6434 led to the induction of TLR tolerance, and no anti-tumor response was observed. We show that TLR7 tolerance was independent of type I interferon (IFN) negative feedback because induction of TLR7 tolerance was also observed in IFN-α/ß receptor knockout mice treated with DSR-6434. Moreover, our data demonstrate that treatment of bone marrow-derived plasmacytoid dendritic cells (BM-pDC) with DSR-6434 led to downregulation of TLR7 expression. From our data, dose scheduling of systemically administered TLR7 agonists can impact on anti-tumor activity through the induction of TLR tolerance. Furthermore, TLR7 expression on pDC may be a useful biomarker of TLR7 tolerance and aid in the optimization of dosing schedules involving systemically administered TLR7 agonists.
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Adenina/análogos & derivados , Carcinoma de Células Renales/inmunología , Glicoproteínas de Membrana/metabolismo , Receptor Toll-Like 7/metabolismo , Adenina/administración & dosificación , Adenina/farmacología , Animales , Antígenos de Neoplasias/inmunología , Línea Celular Tumoral , Protocolos Clínicos , Citotoxicidad Inmunológica , Humanos , Tolerancia Inmunológica , Inmunidad Innata , Interferón Tipo I/metabolismo , Glicoproteínas de Membrana/agonistas , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Neoplasias Experimentales , Transducción de Señal , Receptor Toll-Like 7/agonistasRESUMEN
We have previously reported a series of cyclopropyl urea derivatives as potent orally available soluble epoxide hydrolase (sEH) inhibitors. Here, we designed and synthesized three substituted cyclopropane derivatives that occupy all available pockets of sEH catalytic domain. Compound 14 with a diphenyl substituted cyclopropyl moiety showed good sEH inhibitory activity. Co-crystal structure of this compound and human sEH hydrolase catalytic domain revealed enzyme pockets occupied by the phenoxypiperidine part and the diphenyl cyclopropyl moiety. Furthermore, investigation of the phenoxypiperidine part of compound 14 resulted in the discovery of compound 19, which showed potent sEH inhibitory activity (sub-nM sEH IC50 values).
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Epóxido Hidrolasas/antagonistas & inhibidores , Urea/análogos & derivados , Animales , Sitios de Unión , Dominio Catalítico , Cristalografía por Rayos X , Ciclopropanos/química , Epóxido Hidrolasas/metabolismo , Humanos , Simulación de Dinámica Molecular , Unión Proteica , Ratas , Relación Estructura-Actividad , Urea/síntesis química , Urea/metabolismoRESUMEN
Purpose: To determine the first line of infertility treatment for managing patients with unilateral or bilateral ovarian endometriomas. Methods: We evaluated pregnancy outcome in patients who had received ovarian surgery for unilateral (Group U, n = 47) or bilateral endometriomas (Group B, n = 38) and aspiration with or without alcohol fixation for unilateral (Group u, n = 37) or bilateral endometriomas (Group b, n = 22). Subsequently, 64 of these women, excluding 29 dropouts, underwent assisted reproductive technology. We compared the clinical pregnancy rates of the four groups. Results: The cumulative pregnancy rate after operation of Group B (18%) was significantly lower than that of a cyst-free control group (n = 143; 44%) and Group U (43%). Group B had fewer oocytes fertilized during ART than did Group b (P < 0.005) and fewer blastocysts available for transfer (P < 0.005). The cumulative pregnancy rate of Group B was also lower than in Group b (P = 0.052). Conclusions: Pregnancy outcomes of Group B were not better than for Group b. Therefore, encouraging such women to move directly to ART might help avoid ovarian damage and improve their ability to achieve a pregnancy.
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Aim: To evaluate the efficacy of cryopreservation of all blastocysts for future transfers in stimulated cycles. Methods: We carried out fresh blastocyst transfer cycles on day 5 (n = 290) or day 6 (n = 119) and thawed blastocyst transfer cycles that were frozen on day 5 (n = 136), day 6 (n = 71) or day 6 electively (n = 21). We retrospectively compared the clinical outcome of fresh blastocyst transfers with thawed blastocyst transfers according to the day of blastocyst transfer or freezing. Results: The clinical implantation rates in women with stimulated cycles were significantly higher after the transfer of thawed blastocysts compared with the transfer of fresh blastocysts (day 5, P < 0.0005; day 6, P < 0.00005). Although the implantation rate of fresh day 6 transfer cycles was lower than that of elective day 6 frozen-thawed cycles, this difference was not statistically significant (P = 0.17). Conclusions: Thawed blastocysts demonstrated a better potential for implantation when compared with fresh blastocysts in stimulated cycles. We concluded that elective cryopreservation of all blastocysts on day 5 is an effective option to improve the clinical outcome in stimulated cycles. Additionally, with cryopreservation of all day 6 blastocysts, the implantation rates of first embryo transfers may increase by allowing the best-quality blastocysts to be transferred in thawed cycles. (Reprod Med Biol 2008; 7: 75-83).
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Aim: To assess the appropriateness of assisted hatching using long zona dissection of human frozen-thawed blastocysts at the time of warming, especially in women over 35 years of age or with repeated implantation failures. Methods: Of 177 frozen-thawed blastocyst transfer cycles, 89 control cycles had an intact zona and 88 cycles had assisted hatching using long zona dissection of human thawed blastocyst at the time of warming. These two groups were further subdivided by age to a total of four subgroups: ≤34 years (assisted hatching, n = 39; controls, n = 39) and ≥35 years (assisted hatching, n = 49; controls, n = 50). Twenty-seven cycles in the control group and 28 cycles in the assisted-hatching group had repeated implantation failures. The clinical and ongoing pregnancy rates and the implantation rate between the two groups were analyzed retrospectively. Results: The clinical pregnancy and implantation rates in women ≤34 years were significantly higher after the application of assisted hatching compared with the control group (87.2% and 71.2%vs 56.4% and 46.6%, P < 0.001). The clinical pregnancy and implantation rates of women with repeated implantation failures were higher after the application of assisted hatching compared with the control group (64.3% and 46.3%vs 48.1% and 34.1%), but this difference was not statistically significant. Conclusions: Routine assisted hatching using long zona dissection at the time of warming on frozen-thawed blastocysts is a safe and easy method to perform and is extremely beneficial for increasing the pregnancy rate in young women ≤34 years of age, but not in women ≥35 years of age. Despite increased pregnancy and implantation rates in patients with repeated implantation failures, statistical significance was not achieved. (Reprod Med Biol 2007; 6: 211-218).
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Ischemia-reperfusion injury occurs in acute myocardial infarction, cardiopulmonary bypass surgery, and heart transplantation. However the precise mechanisms still remain unclear. In order to identify proteins that are involved in ischemia-reperfusion injury, we compared precipitated 100,000g fractions of normal, ischemic, and ischemic-reperfused rat hearts using two-dimensional (2D) difference gel electrophoresis (2D-DIGE). 2D-DIGE is reliable method to define quantitative protein differences, especially when subtle protein changes are under investigation. In this study, six spots that changed more than twofold and two additional spots related to these spots were detected. Five of the spots were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry as protein disulfide isomerase, one as 60 kDa heat-shock protein, and two as elongation factor Tu.
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Electroforesis en Gel Bidimensional/métodos , Miocardio/metabolismo , Proteoma/análisis , Daño por Reperfusión/metabolismo , Animales , Chaperonina 60/análisis , Electroforesis en Gel Bidimensional/instrumentación , Infarto del Miocardio/metabolismo , Isquemia Miocárdica/metabolismo , Miocardio/patología , Factor Tu de Elongación Peptídica/análisis , Proteína Disulfuro Isomerasas/análisis , Ratas , Reproducibilidad de los ResultadosRESUMEN
The effects of SM-197378, 2-[[[amino(imino)methyl]amino]carbonyl]-1-methyl-4-trifluoromethyl-1H-indol-7-yl=hydrogen=sulfate monohydrate, a novel potent Na+/H+exchange inhibitor, on heart injury were studied using a rabbit model involving 30 min of myocardial ischemia and 5 h of reperfusion. Intravenous administration of SM-197378 before ischemia reduced the infarct size by approximately 30-50% in a dose-dependent manner. This anti-necrotic effect was achieved without significant hemodynamic changes. Moreover, administration of SM-197378 before reperfusion also resulted in a significant, approximately 30-40%, reduction in the infarct size. The anti-necrotic effect of pre-ischemic bolus treatment with SM-197378 was compared with that of nicorandil, a K+channel opener with nitrate-like activity, and ischemic preconditioning. With 30 and 60 min of ischemia, the anti-necrotic effects of SM-197378 and ischemic preconditioning were similar and superior to that of nicorandil. With 90 min of ischemia, the anti-necrotic effect of SM-197378 was superior to that of ischemic preconditioning.
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Indoles/farmacología , Infarto del Miocardio/prevención & control , Intercambiadores de Sodio-Hidrógeno/antagonistas & inhibidores , Animales , Presión Sanguínea/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Corazón/efectos de los fármacos , Corazón/fisiopatología , Frecuencia Cardíaca/efectos de los fármacos , Precondicionamiento Isquémico , Masculino , Infarto del Miocardio/etiología , Infarto del Miocardio/patología , Isquemia Miocárdica/complicaciones , Daño por Reperfusión Miocárdica/complicaciones , Miocardio/patología , Nicorandil/farmacología , Conejos , Factores de Tiempo , Vasodilatadores/farmacologíaRESUMEN
The effects of 3-[2-({[amino(imino)methyl]amino}carbonyl)-4-chloro-1H-indol-1-yl]-1-propanesulphonic acid monohydrate (SM-198110), a novel potent Na+/H+ exchange inhibitor, and cariporide (Hoe642), another Na+/H+ exchange inhibitor, were studied in a myocardial ischaemia and reperfusion injury model. Anaesthetized rabbits were subjected to occlusion of the coronary artery for 30 min followed by reperfusion for 5 h. SM-198110 or cariporide was administered before ischaemia and before reperfusion. We also assessed the anti-necrotic effect of SM-198110 when given before reperfusion, both alone and together with glibenclamide, a K(ATP) channel blocker, 5-hydroxydecanoate (5-HD), a mitochondrial K(ATP) channel-selective blocker and 8-(p-sulphophenyl)-theophylline (8-SPT), an adenosine receptor blocker. The infarct size was reduced dose-dependently by i.v. administration of SM-198110 before ischaemia, with a significant reduction in serum creatine phosphokinase activity. Infarct sizes, normalized to the size of the area-at-risk (means+/-SE) were: vehicle 56.6+/-3.7%; low-dose SM-198110 39.2+/-6.3%; mid-dose 32.8+/-7.4% (P < 0.05); high-dose 22.1+/-6.7% (P < 0.01). This anti-necrotic effect of SM-198110 was achieved without significant haemodynamic changes. Cariporide given before ischaemia also reduced infarct size significantly and dose-dependently. SM-198110 administered before reperfusion also resulted in a dose-dependent reduction in the infarct size. Infarct sizes were: vehicle 56.6+/-3.7%; low-dose SM-198110 44.5+/-5.7%; mid-dose 36.3+/-6.6% (P < 0.01); high-dose 34.7+/-3.8% (P < 0.01). In contrast, cariporide given before reperfusion did not reduce infarct sizes significantly. The anti-necrotic effect of SM-198110 was observed even when given 10 min after the beginning of reperfusion. Glibenclamide and 5-HD abolished the anti-necrotic effect of treatment before reperfusion with SM-198110. However, the co-administration of 8-SPT with SM-198110 did not affect infarct size. These results suggest that, in addition to Na+/H+ exchange inhibition, mitochondrial and/or sarcolemmal K(ATP) channels contribute to the anti-necrotic effect of SM-198110 when the latter is given before reperfusion.
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Ácidos Alcanesulfónicos/uso terapéutico , Guanidinas/uso terapéutico , Precondicionamiento Isquémico Miocárdico/métodos , Infarto del Miocardio/prevención & control , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Fosfoproteínas/antagonistas & inhibidores , Sulfonas/uso terapéutico , Animales , Presión Sanguínea/efectos de los fármacos , Creatina Quinasa/sangre , Frecuencia Cardíaca/efectos de los fármacos , Masculino , Infarto del Miocardio/etiología , Infarto del Miocardio/patología , Necrosis , Conejos , Intercambiadores de Sodio-HidrógenoRESUMEN
The protective effects of the Na+-H+ exchange (NHE) inhibitors SM-198110 (2-[[(aminoiminomethyl) amino] carbonyl]-4-chloro-1H-indole-1-propanesulfonic acid monohydrate) and SM-197378 (N-(aminoiminomethyl)-1-methyl-7-(sulfooxy)-4-(trifluoromethyl)-1H-indole-2-carboxamide monohydrate) were investigated in perfused Langendorff guinea-pig hearts subjected to ischemia (40 min) and reperfusion (40 min). The recovery of left ventricular developed pressure (LVDP) from ischemia by reperfusion was 39.0% in the control, while in the hearts pretreated with SM-198110 or SM-197378 (10(-7) M), it was about 100%. The ATP level, monitored simultaneously by (31)P-nuclear magnetic resonance spectrometry, was already higher than the control value at the end of the ischemic period, and the elevation in Na+ or Ca2+ fluorometric signals induced during ischemia was suppressed. In post-treated hearts, the LVDP recovery rate was significantly higher with SM-198110 than with SM-197378. By in vitro electron paramagnetic resonance spectrometry, SM-197378 was found to directly quench the active oxygen radical, whereas SM-198110 had no effect. Numbers of apoptotic cardiomyocytes after ischemia (1 h) followed by reperfusion (5 h) were significantly lower in SM-197378-treated than in SM-198110-treated hearts, consistent with the level of activity of caspase-3. These results suggest that the antioxidant effects of NHE inhibitors have an important role in apoptosis during ischemia-reperfusion, but apoptosis is not a major manifestation of cardiac function during postischemic recovery, and that NHE-sensitive mechanisms of reperfusion injury promote both necrotic and apoptotic processes death.
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Apoptosis/efectos de los fármacos , Corazón/efectos de los fármacos , Daño por Reperfusión Miocárdica/patología , Daño por Reperfusión Miocárdica/prevención & control , Intercambiadores de Sodio-Hidrógeno/antagonistas & inhibidores , Animales , Presión Sanguínea/efectos de los fármacos , Caspasa 3 , Caspasas/fisiología , Proteínas de Transporte de Catión/biosíntesis , Proteínas de Transporte de Catión/genética , Separación Celular , Circulación Coronaria/efectos de los fármacos , Citosol/metabolismo , Espectroscopía de Resonancia por Spin del Electrón , Activación Enzimática/efectos de los fármacos , Femenino , Fluorometría , Cobayas , Concentración de Iones de Hidrógeno , Radical Hidroxilo/metabolismo , Etiquetado Corte-Fin in Situ , Técnicas In Vitro , Espectroscopía de Resonancia Magnética , Masculino , Proteínas de la Membrana/biosíntesis , Proteínas de la Membrana/genética , Mitocondrias Cardíacas/efectos de los fármacos , Mitocondrias Cardíacas/metabolismo , Contracción Miocárdica/efectos de los fármacos , Miocardio/patología , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/patología , Ratas , Ratas Sprague-Dawley , Intercambiador de Sodio-Calcio/efectos de los fármacos , Intercambiador 1 de Sodio-Hidrógeno , Intercambiadores de Sodio-Hidrógeno/biosíntesis , Intercambiadores de Sodio-Hidrógeno/genética , Superóxidos/metabolismoRESUMEN
Ischemia-reperfusion injury is a major complication occurring in acute myocardial infarction, cardiopulmonary bypass surgery, and heart transplantation. The aim of this study was to identify proteins that were involved in ischemia-reperfusion injury using fluorescence two-dimensional difference gel electrophoresis. We compared the 100,000 x g precipitate fractions of normal, ischemic and ischemia-reperfused rat hearts and detected six spots which changed more than two-fold in expression level and two additional spots related to these spots. Using peptide mass fingerprinting by matrix-assisted laser desorption/ionization-time of flight mass spectrometry, we identified five of these spots as protein disulfide isomerase A3 (PDA3), one as 60 kDa heat shock protein (HSP60) and two as elongation factor Tu (EF-Tu). HSP60 was increased during ischemia and decreased to normal expression level after reperfusion. EF-Tu was increased in ischemia but not decreased by reperfusion. We also found that several protein spots of PDA3 shifted towards a higher isoelectric point in ischemia and ischemia-reperfusion. Our data strongly suggested that PDA3 underwent dephosphorylation during ischemia and reperfusion and serine 343 of PDA3 was one of the phosphorylation sites.
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Electroforesis en Gel Bidimensional/métodos , Isquemia , Microscopía Fluorescente/métodos , Miocardio/metabolismo , Proteoma , Daño por Reperfusión/patología , Animales , Sitios de Unión , Carbocianinas/química , Chaperonina 60/metabolismo , Bases de Datos como Asunto , Colorantes Fluorescentes/farmacología , Procesamiento de Imagen Asistido por Computador , Masculino , Factor Tu de Elongación Peptídica/metabolismo , Fosforilación , Proteína Disulfuro Isomerasas/química , Ratas , Ratas Wistar , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Fracciones SubcelularesRESUMEN
OBJECTIVE: To investigate anthropometric measures that closely correlate with symptomatic relief of osteoarthritis (OA) of the knee in response to lateral wedged insole use. METHODS: Seventy-one patients with medial compartment knee OA were treated with insoles with subtalar strapping or insoles with talonavicular strapping. Randomization was performed according to birth date. The following variables were evaluated: age, disease duration, Kellgren-Lawrence radiographic stage, body mass index, percent body fat, waist to hip ratio, lower extremity lean body mass (L-LBM) per body weight, and radiographic femorotibial angle at baseline. The trial lasted 8 weeks. The correlation between each variable and the remission score (delta score) using the Lequesne index of severity was analyzed. RESULTS: In the subtalar strapping group (n = 34), delta score of knee OA was more strongly associated with age (p = 0.004, r = 0.48) than other variables. A significant correlation was also observed between L-LBM per body weight and delta score (p = 0.041, r = -0.36) in the subtalar strapping group. No other variables significantly correlated with the delta score in the subtalar strapping groups. No variable significantly correlated with the ? score in the talonavicular strapping group (n = 37). CONCLUSION: We previously reported that use of insoles with subtalar strapping leads to valgus realignment of the femorotibial angle in patients with knee OA with varus deformity, and it may have a similar therapeutic effect to that of high tibial osteotomy. These data suggest that the insole with subtalar strapping is more efficacious for younger patients and those with a higher L-LBM per body weight, and less efficacious for older patients with sarcopenia.
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Composición Corporal , Aparatos Ortopédicos , Osteoartritis de la Rodilla/fisiopatología , Osteoartritis de la Rodilla/terapia , Factores de Edad , Anciano , Femenino , Fémur/diagnóstico por imagen , Pie , Humanos , Persona de Mediana Edad , Osteoartritis de la Rodilla/diagnóstico por imagen , Satisfacción del Paciente , Radiografía , Tibia/diagnóstico por imagenRESUMEN
The aim of this study was to investigate whether a selective Na+/H+ exchange inhibitor, SM-20550, can modulate the mitochondrial respiratory function and mitochondrial Ca2+ content in isolated rat hearts subjected to 40 min of ischemia and 20 min of reperfusion. SM-20550 (10, 100 nM) was administered for 5 min prior to ischemia and for 20 min during the reperfusion period. At 20 min after reperfusion, treatment with SM-20550 (10, 100 nM) improved the recovery of left ventricular developed pressure and suppressed the rise in left ventricular end-diastolic pressure. Mitochondrial function, assessed by the state 3 oxygen respiration rate, respiratory control index, and oxidative phosphorylation rate, was significantly impaired after ischemia/reperfusion. Administration with SM-20550 (10, 100 nM) attenuated the impaired mitochondrial function, improving the state 3 respiration rate, respiratory control index, and oxidative phosphorylation rate. The mitochondrial Ca2+ content was significantly increased after ischemia/reperfusion but was suppressed by treatment with SM-20550 (10, 100 nM). A significant linear correlation was observed between the respiratory control index and mitochondrial Ca2+ content in the ischemic/reperfused hearts. In conclusion, SM-20550 improved the postischemic recovery of left ventricular function and concurrently protected mitochondrial function mediated by preventing mitochondrial Ca2+ overload.
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Amidinas/farmacología , Calcio/metabolismo , Indoles/farmacología , Mitocondrias Cardíacas/efectos de los fármacos , Mitocondrias Cardíacas/metabolismo , Isquemia Miocárdica/fisiopatología , Daño por Reperfusión Miocárdica/fisiopatología , Intercambiadores de Sodio-Hidrógeno/antagonistas & inhibidores , Animales , Técnicas In Vitro , Masculino , Miocardio/metabolismo , Miocardio/patología , Fosforilación Oxidativa/efectos de los fármacos , Consumo de Oxígeno/efectos de los fármacos , Ratas , Ratas Wistar , Función Ventricular Izquierda/efectos de los fármacosRESUMEN
The effect of a novel potent Na(+)/H(+) exchange inhibitor, SM-20550 [N-(aminoiminomethyl)-1,4-dimethyl-1H-indole-2-carboxamide methanesulfonic acid], on survival after myocardial infarction was studied. Anesthetized rats underwent occlusion of the coronary artery (30 min) followed by reperfusion (14 days). SM-20550 was administered intravenously before ischemia (1-day treatment group) or before ischemia and on the 2 days following (3-day treatment group). The infarct size was significantly reduced on the 14th day after surgery by approximately 17 and 20% in 1- and 3-day treatment groups, respectively. The survival rate on day 14 was significantly enhanced in both treatment groups (96%) compared with the vehicle-treated control group (70%). These results suggest that SM-20550 improved survival after myocardial infarction, at least due to its antinecrotic effect.