RESUMO
Piscirickettsia salmonis is the most important health problem facing Chilean Aquaculture. Previous reports suggest that P. salmonis can survive in salmonid macrophages by interfering with the host immune response. However, the relevant aspects of the molecular pathogenesis of P. salmonis have been poorly characterized. In this work, we evaluated the transcriptomic changes in macrophage-like cell line SHK-1 infected with P. salmonis at 24- and 48-hours post-infection (hpi) and generated network models of the macrophage response to the infection using co-expression analysis and regulatory transcription factor-target gene information. Transcriptomic analysis showed that 635 genes were differentially expressed after 24- and/or 48-hpi. The pattern of expression of these genes was analyzed by weighted co-expression network analysis (WGCNA), which classified genes into 4 modules of expression, comprising early responses to the bacterium. Induced genes included genes involved in metabolism and cell differentiation, intracellular transportation, and cytoskeleton reorganization, while repressed genes included genes involved in extracellular matrix organization and RNA metabolism. To understand how these expression changes are orchestrated and to pinpoint relevant transcription factors (TFs) controlling the response, we established a curated database of TF-target gene regulatory interactions in Salmo salar, SalSaDB. Using this resource, together with co-expression module data, we generated infection context-specific networks that were analyzed to determine highly connected TF nodes. We found that the most connected TF of the 24- and 48-hpi response networks is KLF17, an ortholog of the KLF4 TF involved in the polarization of macrophages to an M2-phenotype in mammals. Interestingly, while KLF17 is induced by P. salmonis infection, other TFs, such as NOTCH3 and NFATC1, whose orthologs in mammals are related to M1-like macrophages, are repressed. In sum, our results suggest the induction of early regulatory events associated with an M2-like phenotype of macrophages that drives effectors related to the lysosome, RNA metabolism, cytoskeleton organization, and extracellular matrix remodeling. Moreover, the M1-like response seems delayed in generating an effective response, suggesting a polarization towards M2-like macrophages that allows the survival of P. salmonis. This work also contributes to SalSaDB, a curated database of TF-target gene interactions that is freely available for the Atlantic salmon community.
Assuntos
Salmo salar , Animais , Salmo salar/genética , Perfilação da Expressão Gênica , Macrófagos/metabolismo , Fatores de Transcrição/metabolismo , RNA/metabolismo , MamíferosRESUMO
BACKGROUND AND PURPOSE: Synchronous bilateral breast cancer (SBBC) accounts for 1-3.5% of breast cancer patients. The aim of this study was to evaluate dosimetric issues, clinical outcomes, and acute toxicities for SBBC patients receiving synchronous bilateral hypofractionated radiotherapy (SBHRT) and to compare them with patients treated with synchronous bilateral normofractionated RT schedule (SBNRT). MATERIALS AND METHODS: From April 2016 to March 2020, 39 SBBC patients were referred to our institution. Patients were divided according to their prescription dose: Group A: 50 Gy/25fx (fractions), B: 60-64 Gy/25fx, C: 40.05 Gy/15fx; D: 48 Gy/15fx. Toxicity was evaluated using Common Terminology Criteria for Adverse Events (CTCAE)v.5.0. RESULTS: 34 patients were finally evaluated. Median follow-up was 24 months for NF schedule and 9 months for HF schedule. In the HF schedule, no acute side-effects > G2 were observed and no dermatitis was reported in 6th month´s assessments. 95% of patients have no evidence of disease and only 1 patient presented local relapse in the first mammography after RT. No distant failures or deaths were observed. Regarding dosimetric issues, the inter-patient average Dmean for the heart was: Group A: 5.0 Gy (4.6-5.5), Group B: 4.4 Gy (4.1-5.4), Group C: 4.8 Gy (4.5-5.1) and Group D: 5.3 Gy (4.4-5.6). For the lungs, the inter-patient average Dmean was: Group A: 10.8 Gy (9.8-12.2), Group B: 11.5 Gy (11.3-12), Group C: 9.8 Gy (9.3-10.5) and Group D: 10.5 Gy (10-11.3). CONCLUSIONS: This is the first study reporting the safety, feasibility, and tolerability of 40.05 Gy/15fx over 3 weeks for the treatment of SBBC patients. Further study with larger accrual is mandatory.
Assuntos
Neoplasias da Mama/radioterapia , Neoplasias Primárias Múltiplas/radioterapia , Hipofracionamento da Dose de Radiação , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/cirurgia , Estudos de Viabilidade , Feminino , Seguimentos , Coração/efeitos da radiação , Humanos , Pulmão/efeitos da radiação , Neoplasias Primárias Múltiplas/diagnóstico por imagem , Neoplasias Primárias Múltiplas/cirurgia , Órgãos em Risco/efeitos da radiação , Estudos Prospectivos , Radioterapia de Intensidade Modulada/efeitos adversos , Fatores de TempoRESUMO
PURPOSE: To study the control graphs applicability for the geometric uncertainties of VMAT treatments in prostate cancer patients, and their use to verify the hypothesis of the data obtained randomness, to apply the margins of Van Herk expression. MATERIAL AND METHODS: During the first 5 days of treatment, and then once a week, a Kv CBCT was performed, compared with the simulation CT and adjusted the displacements, to determine the inter-fraction errors. Immediately after radiation therapy, another CBCT was performed (for intra-fraction errors). With these data, the X, R position control charts have been made. The patients, not maintained the deviations within the charts control limits, were called "anomalies". Then, we compared the deviations and margins calculated with the van Herk expression for all patients and for those without anomalies. RESULTS: The margins determined show appreciable differences if there were calculated for the total set of patients or for the set of them without anomalies in the control charts. For the overall set of patients, the lateral, longitudinal, and vertical margins were 0.45 cm, 0.52 cm, 0.56 cm, while for the set of patients without anomalies were 0.29 cm, 0.35 cm, and 0.38 cm. CONCLUSIONS: The use of control charts allows tracking geometric deviations both inter and intra-fraction, variability real-time control and to detect situations in which it can change for non-random reasons, and require immediate investigation. Maintaining geometric deviations in the control state decreases the margins needed to administer a high dose to CTV in a high percentage of cancer prostate patients.
Assuntos
Neoplasias da Próstata/patologia , Neoplasias da Próstata/radioterapia , Dosagem Radioterapêutica , Radioterapia de Intensidade Modulada/estatística & dados numéricos , Incerteza , Humanos , Masculino , Estudos ProspectivosRESUMO
Piscirickettsia salmonis, an aggressive intracellular pathogen, is the etiological agent of salmonid rickettsial septicemia (SRS). This is a chronic multisystemic disease that generates high mortalities and large losses in Chilean salmon farming, threatening the sustainability of the salmon industry. Previous reports suggest that P. salmonis is able to survive and replicate in salmonid macrophages, inducing an anti-inflammatory environment and a limited lysosomal response that may be associated with host immune evasion mechanisms favoring bacterial survival. Current control and prophylaxis strategies against P. salmonis (based on the use of antibiotics and vaccines) have not had the expected success against infection. This makes it urgent to unravel the host-pathogen interaction to develop more effective therapeutic strategies. In this study, we evaluated the effect of treatment with IgM-beads on lysosomal activity in Atlantic salmon macrophage-enriched cell cultures infected with P. salmonis by analyzing the lysosomal pH and proteolytic ability through confocal microscopy. The impact of IgM-beads on cytotoxicity induced by P. salmonis in infected cells was evaluated by quantification of cell lysis through release of Lactate Dehydrogenase (LDH) activity. Bacterial load was determined by quantification of 16S rDNA copy number by qPCR, and counting of colony-forming units (CFU) present in the extracellular and intracellular environment. Our results suggest that stimulation with antibodies promotes lysosomal activity by lowering lysosomal pH and increasing the proteolytic activity within this organelle. Additionally, incubation with IgM-beads elicits a decrease in bacterial-induced cytotoxicity in infected Atlantic salmon macrophages and reduces the bacterial load. Overall, our results suggest that stimulation of cells infected by P. salmonis with IgM-beads reverses the modulation of the lysosomal activity induced by bacterial infection, promoting macrophage survival and bacterial elimination. This work represents a new important evidence to understand the bacterial evasion mechanisms established by P. salmonis and contribute to the development of new effective therapeutic strategies against SRS.
Assuntos
Anticorpos Antibacterianos/imunologia , Doenças dos Peixes/imunologia , Lisossomos/imunologia , Macrófagos/imunologia , Piscirickettsia/imunologia , Infecções por Piscirickettsiaceae/imunologia , Salmão/imunologia , Animais , Doenças dos Peixes/microbiologia , Lisossomos/microbiologia , Macrófagos/microbiologia , Infecções por Piscirickettsiaceae/veterinária , Salmão/microbiologiaRESUMO
BACKGROUND: There is an emerging field to put into practice new strategies for developing molecules with antimicrobial properties. In this line, several metals and metalloids are currently being used for these purposes, although their cellular effect(s) or target(s) in a particular organism are still unknown. Here we aimed to investigate and analyze Au3+ toxicity through a combination of biochemical and molecular approaches. RESULTS: We found that Au3+ triggers a major oxidative unbalance in Escherichia coli, characterized by decreased intracellular thiol levels, increased superoxide concentration, as well as by an augmented production of the antioxidant enzymes superoxide dismutase and catalase. Because ROS production is, in some cases, associated with metal reduction and the concomitant generation of gold-containing nanostructures (AuNS), this possibility was evaluated in vivo and in vitro. CONCLUSIONS: Au3+ is toxic for E. coli because it triggers an unbalance of the bacterium's oxidative status. This was demonstrated by using oxidative stress dyes and antioxidant chemicals as well as gene reporters, RSH concentrations and AuNS generation.
Assuntos
Escherichia coli/efeitos dos fármacos , Ouro/toxicidade , Nanopartículas Metálicas/toxicidade , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacosRESUMO
The posterior arch defects of the cervical spine are rare, and they arise out of deviations of the normal intrauterine development of the spine (4-8 weeks of development). The defects range from a cleft to the total agenesis of the posterior arch, with a reported prevalence of 4% and 0.15%, respectively. The pedicle agenesis is most frequently found in C6. A diagnosis is usually made after a traumatic incident in a previously asymptomatic patient. 35% of a patient's current symptoms are associated with instability or translation of the impaired vertebral segments like headaches, chronic pain, and neurological impairment. The clinical and radiological findings of a patient with an unusual and complex cervical spine malformation are reported. These are rare entities and seldom require surgical treatment. It is imperative for spine surgeons to be aware of these anatomical abnormalities to avoid misinterpretation and thus inappropriate treatment, particularly in acute trauma patients.
Assuntos
Vértebras Cervicais/anormalidades , Pescoço/anormalidades , Fusão Vertebral , Placas Ósseas , Vértebras Cervicais/diagnóstico por imagem , Vértebras Cervicais/cirurgia , Feminino , Humanos , Imageamento por Ressonância Magnética , Pescoço/diagnóstico por imagem , Pescoço/cirurgia , Tomografia Computadorizada por Raios X , Resultado do Tratamento , Adulto JovemRESUMO
PURPOSE: We performed a cross-sectional study of neurocognitive function in non-brain cancer patients treated with long-term bevacizumab. METHODS/PATIENTS: From 2015 to 2017, we included patients with different types of cancer treated with bevacizumab with or without chemotherapy (BEV; N = 20) or only chemotherapy (ChT; N = 19) for at least 34 weeks, patients who received non-brain radiotherapy (RxT; N = 19), and healthy controls (HC; N = 19) were assessed once at week 34 of treatment (BEV and ChT) or at completion of radiotherapy. Neurocognition was evaluated with the Hopkins Verbal Learning Test-Revised (HVLT-R) total and delayed recall, the Trail Making Test A and B, and the Controlled Oral Word Association Test in the four groups. Non-parametric tests were used to assess differences between groups. RESULTS: The BEV, ChT, and RxT groups scored significantly lower than the HC group on all tests and especially on the HVLT-R total recall. In no case were the mean scores of the BEV group significantly lower than those of the ChT or RxT groups. CONCLUSIONS: Neurocognitive impairment was seen even in patients treated with local non-brain radiotherapy. Treatment with bevacizumab for a long period of time does not seem to worsen neurocognitive function to a greater extent than chemotherapy.
Assuntos
Antineoplásicos Imunológicos/uso terapêutico , Bevacizumab/uso terapêutico , Neoplasias/tratamento farmacológico , Transtornos Neurocognitivos/diagnóstico , Antineoplásicos Imunológicos/efeitos adversos , Bevacizumab/efeitos adversos , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias/radioterapia , Transtornos Neurocognitivos/etiologia , Testes NeuropsicológicosRESUMO
BACKGROUND: There Is an emerging field to put Into practice new strategies for developing molecules with antimicrobial properties. In this line, several metals and metalloids are currently being used for these purposes, although their cellular effect(s) or target(s) in a particular organism are still unknown. Here we aimed to investigate and analyze Au3+ toxicity through a combination of biochemical and molecular approaches. RESULTS: We found that Au3+ triggers a major oxidative unbalance in Escherichia coli, characterized by decreased intracellular thiol levels, increased superoxide concentration, as well as by an augmented production of the antioxidant enzymes superoxide dismutase and catalase. Because ROS production is, in some cases, associated with metal reduction and the concomitant generation of gold-containing nanostructures (AuNS), this possibility was evaluated in vivo and in vitro. CONCLUSIONS: Au3+ is toxic for E. coli because it triggers an unbalance of the bacterium's oxidative status. This was demonstrated by using oxidative stress dyes and antioxidant chemicals as well as gene reporters, RSH concentrations and AuNS generation.
Assuntos
Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Nanopartículas Metálicas/toxicidade , Ouro/toxicidadeRESUMO
To enhance our understanding of the control of archaeal carbon central metabolism, a detailed analysis of the regulation mechanisms of both fructose1,6-bisphosphatase (FruBPase) and ADP-phosphofructokinase-1 (ADP-PFK1) was carried out in the methanogen Methanosarcina acetivorans. No correlations were found among the transcript levels of the MA_1152 and MA_3563 (frubpase type II and pfk1) genes, the FruBPase and ADP-PFK1 activities, and their protein contents. The kinetics of the recombinant FruBPase II and ADP-PFK1 were hyperbolic and showed simple mixed-type inhibition by AMP and ATP, respectively. Under physiological metabolite concentrations, the FruBPase II and ADP-PFK1 activities were strongly modulated by their inhibitors. To assess whether these enzymes were also regulated by a phosphorylation/dephosphorylation process, the recombinant enzymes and cytosolic-enriched fractions were incubated in the presence of commercial protein phosphatase or protein kinase. De-phosphorylation of ADP-PFK1 slightly decreased its activity (i.e. Vmax) and did not change its kinetic parameters and oligomeric state. Thus, the data indicated a predominant metabolic regulation of both FruBPase and ADP-PFK1 activities by adenine nucleotides and suggested high degrees of control on the respective pathway fluxes.
Assuntos
Proteínas Arqueais/metabolismo , Frutose-Bifosfatase/metabolismo , Methanosarcina/metabolismo , Fosfofrutoquinase-1/metabolismo , Difosfato de Adenosina/metabolismo , Monofosfato de Adenosina/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Proteínas Arqueais/genética , Proteínas Arqueais/isolamento & purificação , Galinhas , Frutose-Bifosfatase/genética , Frutose-Bifosfatase/isolamento & purificação , Frutosefosfatos/metabolismo , Genes Arqueais , Cinética , Methanosarcina/genética , Fosfofrutoquinase-1/genética , Fosfofrutoquinase-1/isolamento & purificação , Fosforilação , Inibidores de Proteínas Quinases/metabolismo , Processamento de Proteína Pós-TraducionalRESUMO
PURPOSE: The aim of the study was to evaluate the effectiveness of fentanyl pectin nasal spray (FPNS) in controlling procedural breakthrough cancer pain (BTCP) in advanced cancer patients undergoing radiotherapy. MATERIALS AND METHODS: This study involved 62 advanced cancer patients, with well-controlled background pain, who presented BTCP associated to routine radiotherapy procedures, treated with FPNS according to our protocol of administration. The BPE intensity was measured using a visual analog scale (VAS). RESULTS: The BTCP was triggered during the computed tomography simulation (79.3%) or treatment delivery (20.7%). Patients indicated a mean VAS of 8.8 (range 7-10) when attempting the procedure. After 4.5 min (range 2-10) of the first FPNS dose, the majority of patients (85.5%) indicated a VAS of 4.3 (range 2-6). 15.5% of the patients did not respond after 15 min; requiring a second dose. All these patients responded, reporting a mean VAS of 4.2 (range 4-6) after 3.0 min (range 2-5) of the second dose. None of the patients required a third dose, nor reported an AE after the administration of FPNS. CONCLUSIONS: In our knowledge, our study is the one of highest recruitment, and with the fastest response of BTCP treated with FPNS reported in advanced cancer patients undergoing radiotherapy. FPNS has proven to be highly effective in reducing the intensity of procedural BTCP in a very short period of time.
Assuntos
Analgésicos Opioides/administração & dosagem , Dor Irruptiva/tratamento farmacológico , Dor do Câncer/tratamento farmacológico , Fentanila/administração & dosagem , Neoplasias/radioterapia , Dor Processual/tratamento farmacológico , Adulto , Idoso , Idoso de 80 Anos ou mais , Dor Irruptiva/etiologia , Dor do Câncer/etiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Sprays Nasais , Medição da Dor , Dor Processual/complicações , Radioterapia/efeitos adversosRESUMO
PURPOSE: Our aim was investigate whether lymph node uptake is associated with survival and regional relapses, and relapse patterns with respect to the radiotherapy fields in esophageal cancer (EC). MATERIALS AND METHODS: The FDG-PET/CT image datasets of 56 patients were analyzed. All patients underwent definitive or neoadjuvant radio/chemotherapy (RCT). All patients suffering from persistent or recurrent local/regional-only disease after RCT were considered for salvage resection. Patients with adenocarcinoma without metastatic disease were considered for planned resection (usually within 3 months of treatment). RESULTS: Patients with PET-positive lymph nodes before treatment had a worse overall survival and a shorter disease-free survival than those without PET-positive nodes. They also had worse node and metastatic relapse-free survival. N2 patients had statistically significant poorer outcomes than N1-N0 patients and a better survival if the involved nodes were closer to the esophageal tumor. Involved node location by PET/CT also affected global, nodal and metastatic relapses. In addition, an increment of SUVmax value increased relative risk of death and increased relative risk of node and metastatic relapses. The first site of relapse was metastatic recurrence and, second, local recurrence. The most frequent were "in-field" loco/regional recurrence. We observed a relationship between patients classified-N1 and out-field nodal recurrence (p = 0.024), and between patients-N2 and in-field nodal recurrence. The number of PET-positive nodes was an independent significant prognostic predictor for relapse (p < 0.001). CONCLUSION: Our study shows that only FDG-PET/CT can provide prognostic information in EC. Nodal PET/CT uptake influences outcome and relapse location among EC patients.
Assuntos
Adenocarcinoma/mortalidade , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Carcinoma de Células Escamosas/mortalidade , Quimiorradioterapia/mortalidade , Neoplasias Esofágicas/mortalidade , Linfonodos/patologia , Recidiva Local de Neoplasia/mortalidade , Adenocarcinoma/diagnóstico por imagem , Adenocarcinoma/secundário , Adenocarcinoma/terapia , Carcinoma de Células Escamosas/diagnóstico por imagem , Carcinoma de Células Escamosas/secundário , Carcinoma de Células Escamosas/terapia , Neoplasias Esofágicas/diagnóstico por imagem , Neoplasias Esofágicas/patologia , Neoplasias Esofágicas/terapia , Feminino , Seguimentos , Humanos , Linfonodos/metabolismo , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/diagnóstico por imagem , Recidiva Local de Neoplasia/patologia , Recidiva Local de Neoplasia/terapia , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Prognóstico , Compostos Radiofarmacêuticos , Estudos Retrospectivos , Taxa de SobrevidaRESUMO
PURPOSE: Evidence supporting the use of 18F-FDG-PET/CT in the segmentation process of oesophageal cancer for radiotherapy planning is limited. Our aim was to compare the volumes and tumour lengths defined by fused PET/CT vs. CT simulation. MATERIALS AND METHODS: Twenty-nine patients were analyzed. All patients underwent a single PET/CT simulation scan. Two separate GTVs were defined: one based on CT data alone and another based on fused PET/CT data. Volume sizes for both data sets were compared and the spatial overlap was assessed by the Dice similarity coefficient (DSC). RESULTS: The gross tumour volume (GTVtumour) and maximum tumour diameter were greater by PET/CT, and length of primary tumour was greater by CT, but differences were not statistically significant. However, the gross node volume (GTVnode) was significantly greater by PET/CT. The DSC analysis showed excellent agreement for GTVtumour, 0.72, but was very low for GTVnode, 0.25. CONCLUSIONS: Our study shows that the volume definition by PET/CT and CT data differs. CT simulation, without taking into account PET/CT information, might leave cancer-involved nodes out of the radiotherapy-delineated volumes.
Assuntos
Carcinoma de Células Escamosas/radioterapia , Neoplasias Esofágicas/radioterapia , Fluordesoxiglucose F18 , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Planejamento da Radioterapia Assistida por Computador/métodos , Carga Tumoral , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/patologia , Quimiorradioterapia , Neoplasias Esofágicas/diagnóstico , Neoplasias Esofágicas/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Tomografia por Emissão de Pósitrons/métodos , Estudos Retrospectivos , Carga Tumoral/efeitos da radiaçãoRESUMO
Eight new fluorescent biosensors of human calmodulin (hCaM) using Alexa Fluor® 350, 488, 532, and 555 dyes were constructed. These biosensors are thermodynamically stable, functional, and highly sensitive to ligands of the CaM. They resolve the problem of CaM ligands with similar spectroscopic properties to the intrinsic and extrinsic fluorophores of other biosensors previously reported. Additionally, they can be used in studies of protein-protein interaction through Förster resonance energy transfer (FRET). The variation in Tm (range 78.07-81.47 °C; 79.05 to WT) is no larger than two degrees in all cases in regards to CaM WT. The Kds calculated with all biosensors for CPZ and BIMI (a new inhibitor of CaM) are in the range of 0.45-1.86 and 0.69-1.54 µm respectively. All biosensors retain their ability to activate Calcineurin about 70%. Structural models built "in silico" show their possible conformation taking the fluorophores in protein thus we can predict system stability. Finally, these new biosensors represent a biotechnological development applied to an analytical problem, which aims to determine accurately the affinity of inhibitors of CaM without possible interference, to be put forward as possible drugs related to CaM.
Assuntos
Calmodulina/química , Cromonas/química , Inibidores Enzimáticos/química , Transferência Ressonante de Energia de Fluorescência/métodos , Corantes Fluorescentes/química , Modelos Moleculares , HumanosRESUMO
Cottonseed hulls are co-product of agribusiness that can be used in beef cattle rations, decreasing the cost of feed. The aim of this study was to evaluate the effects of different cottonseed hull levels, display and ageing times on visual and sensorial meat acceptability. Longissimus thoracis muscle from 30 crossbred young bulls finished on three high-grain diets (210, 270 or 330 g/kg of cottonseed hulls on dry matter, respectively) were visually evaluated during 10 days of display by 37 appraisers. Tenderness, flavour and overall acceptability from the three diets and three ageing times (1, 7 and 14 days) were evaluated by 120 consumers. On the visual study, time of display (P⩽0.001) was a more significant factor than diet. Cottonseed hull level had no effect on sensorial analyses, with tenderness acceptability improving with ageing time (P⩽0.001). Results indicate the possibility of using the three studied levels of cottonseed without damaging consumer meat acceptability.
Assuntos
Ração Animal/análise , Bovinos/fisiologia , Fibras na Dieta , Carne Vermelha/normas , Animais , Dieta/veterinária , Grão Comestível , Gossypium , Masculino , Sementes , PaladarRESUMO
Introducción: La hipoacusia es una condición prevalente que tiene consecuencias en el desarrollo de los individuos. Cada vez cobran mayor importancia los programas de tamizaje de hipoacusia. Una estrategia involucra la utilización de cuestionarios de tamizaje de hipoacusia. Objetivos: Realizar la adaptación transcultural al español de los instrumentos de tamizaje auditivos ABEL y TEAP. Material y método: Se realizó un estudio de corte transversal de validación de cuestionario en escolares de cuarto básico. Se utilizó la teoría clásica de tests, que incluye validez de criterio, confiabilidad test-retest y consistencia interna. Resultados: Fueron evaluados 100 escolares de San Bernardo. Cincuenta y dos por ciento de la muestra fueron mujeres. El promedio (DE) de edad fue 9,75 (± 0,78) años. Hubo tres casos de hipoacusia bilateral y dos de hipoacusia unilateral. La consistencia interna del test ABEL fue de 0,73. La consistencia interna del test TEAP fue de 0,82. La confiabilidad test-retest del ABEL fue 0,86 y del TEAP fue 0,86. No hubo diferencia entre los puntajes del ABEL ni del TEAP entre niños con hipoacusia y sin hipoacusia. Conclusión: Sería de utilidad utilizar cuestionarios para el tamizaje de hipoacusia en escolares. Estos resultados permiten plantear un programa formal de tamizaje basado en cuestionarios de hipoacusia.
Introduction: Hearinglossis a prevalent condition that has consequences for children's development. As a result, screening programs for hearing loss are of increasing importance. One screening strategy involves the use of questionnaires for people with hearing loss. Aim: Transcultural adaptation to Spanish of the hearing screening in struments ABEL and the TEAP. Material and method: A cross-sectional questionnaire validation study in fourth grade school children was conducted. Classical test theory, including criterion validity, test-retest reliability and internal consistency was used. Results: A total of 100 school children in San Bernardo were evaluated. 52% of the sample were girls. The mean (SD) age was 9.75 (± 0.78) years. There were three cases of bilateral hearing loss and two cases of unilateral hearing loss. The internal consistency of the test was 0.73 for ABEL. The internal consistency was 0.82 for TEAP. The test-retest reliability was 0.86 for ABEL and for TEAP was 0.86. There was no difference between the scores of ABEL or TEAP between the children with hearing loss and those without hearing loss. Conclusions: It would be helpful to use questionnaires to screen for hearing loss in school. These results support a formal screening program based on questionnaires of hearing loss.
Assuntos
Humanos , Masculino , Feminino , Criança , Programas de Rastreamento , Inquéritos e Questionários , Perda Auditiva/epidemiologia , Traduções , Chile , Estudos Transversais , Reprodutibilidade dos TestesRESUMO
Desde hace décadas se sabe que al colocar un electrodo cerca de la ventana redonda de la cóclea es posible registrar potenciales cocleares en respuesta a estímulos sonoros, tales como el potencial microfónico coclear o el potencial de acción compuesto del nervio auditivo. Sin embargo, hace algunos años, se ha descrito que al posicionar un electrodo en el nicho de la ventana redonda en animales de experimentación y en ausencia de estimulación acústica, se obtiene actividad eléctrica aparentemente aleatoria y no desencadenada por estímulo. Esta actividad eléctrica se ha denominado ruido eléctrico de ventana redonda (RWN, por su sigla en inglés Round Window Noise) y representaría la descarga espontánea de fibras del nervio auditivo. La técnica para su registro es similar a la utilizada en la electrococleografía para obtener potenciales cocleares en humanos. Esta señal se distingue del ruido eléctrico no biológico por tener un peak en su poder espectral centrado en torno a los 900 Hz. La amplitud de esta banda de frecuencia se correlaciona con la sensibilidad auditiva en la región basal de la cóclea, entre los 12 a 30 kHz en cobayos y gatos. El RWN se ha estudiado a través de estimulación acústica y bloqueo farmacológico, logrando caracterizar sus propiedades electrofisiológicas para desarrollar modelos de estudio con aplicación clínica.
For decades, it has been known that placing an electrode near the round window niche allows the recording of cochlear potentials in response to auditory stimuli, such as the cochlear microphonic potential and the compound action potential of the auditory nerve. However, some years ago, it was discovered, that by positioning an electrode in the round window niche of experimental animals, and in the absence of acoustic stimulation, apparently random electrical activity is obtained. This electrical activity is called round window electrical noise (RWN) and represents the spontaneous discharge of auditory nerve fibers. The recording technique is similar to that used in electrocochleography in humans. This bioelectrical signal is distinguished from that of non-biological electrical noise, as it has a power spectral peak centered around 900 Hz. The magnitude of this frequency peak is correlated with hearing sensitivity in the basal region of the cochlea, between 12 to 30 kHz in guinea pigs and cats. The RWN has been characterized using sound stimulation or pharmacologic suppression, and its electrophysiological properties could be used to develop models of auditory nerve function with possible clinical application.
Assuntos
Humanos , Janela da Cóclea/fisiologia , Potenciais de Ação/fisiologia , Nervo Coclear/fisiologia , Audiometria de Resposta EvocadaRESUMO
The hippocampus, a medial temporal lobe structure necessary for the formation of spatial memory, is particularly affected by both normal and pathologic aging. In previous studies, we observed a significant age-related increase in dopaminergic neuron loss in the hypothalamus and the substantia nigra of female rats, which becomes more conspicuous at extreme ages. Here, we extend our studies by assessing spatial memory in 4-6 month-old (young), 26-month-old (old) and 29-32-month-old (senile) Sprague-Dawley female rats as well as the age-related histopathological changes in their dorsal hippocampus. Age changes in spatial memory performance were assessed with a modified version of the Barnes maze test. We employed two probe trials (PTs), one and five days after training, respectively, in order to evaluate learning ability as well as short-term and longer-term spatial memory retention. A set of relevant hippocampal cell markers was also quantitated in the animals by means of an unbiased stereological approach. The results revealed that old rats perform better than senile rats in acquisition trials and young rats perform better than both aging groups. However, during short-term PT both aging groups showed a preserved spatial memory while in longer-term PT, spatial memory showed deterioration in both aged groups. Morphological analysis showed a marked decrease (94-97%) in doublecortin neuron number in the dentate gyrus in both aged groups and a reduction in glial fibrillary acidic protein-positive cell number in the stratum radiatum of aging rats. Astroglial process length and branching complexity decreased in aged rats. We conclude that while target-seeking activity and learning ability decrease in aged females, spatial memory only declines in the longer-term tests. The reduction in neuroblast number and astroglial arborescence complexity in the dorsal hippocampus are likely to play a role in the cognitive deficits of aging rats.
Assuntos
Envelhecimento/patologia , Envelhecimento/psicologia , Hipocampo/patologia , Memória Espacial/fisiologia , Animais , Astrócitos/patologia , Cognição/fisiologia , Proteína Duplacortina , Feminino , Neurônios/patologia , Ratos , Ratos Sprague-DawleyRESUMO
PURPOSE: Sorafenib is a multikinase inhibitor that targets the MAPK pathway and is currently used for the treatment of hepatocellular and renal carcinoma. Recently, it has been shown that sorafenib is also cytotoxic to multiple myeloma (MM) cells. Here, we have further analyzed the mechanism of sorafenib-induced death in MM cells. METHODS: Cell death induced by sorafenib in MM cell lines and in plasma cells from MM patients was evaluated by analysis of gene expression by RT-MLPA and quantitative PCR, protein levels and functionality by Western blot and flow cytometry and gene silencing with siRNA. RESULTS: Cell death was characterized by phosphatidylserine exposure, ΔΨm loss, cytochrome c release and caspase activation, hallmarks of apoptosis. DL50 at 24 h ranged from 6 to 10 µM. Ex vivo treatment with 20 µM sorafenib induced apoptosis in around 80 % myeloma cells from six multiple myeloma patients. Sorafenib induced caspase-dependent degradation of Bcl-xL and Mcl-1 proteins, destabilizing the mitochondria and speeding up the development of apoptosis. Sorafenib treatment increased levels of Puma at mRNA and protein level and gene silencing with siRNA confirmed a relevant role for Puma in the induction of apoptosis. Co-treatment with the pan-caspase inhibitor Z-VAD-fmk prevented cell death to a variable degree depending on the cell line. In RPMI 8226 cells, Z-VAD-fmk prevented most of sorafenib-induced death. However, death in MM.1S was only prevented by co-incubation with both Z-VAD-fmk and the RIP1K inhibitor necrostatin-1, indicating that under conditions of inefficient caspase activation, sorafenib induces death by necroptosis. CONCLUSION: Our results demonstrate a key role for Puma in the triggering of sorafenib-induced apoptosis and that this drug can also induce death by necroptosis in multiple myeloma cells.
Assuntos
Antineoplásicos/farmacologia , Proteínas Reguladoras de Apoptose/metabolismo , Apoptose/efeitos dos fármacos , Mitocôndrias/patologia , Mieloma Múltiplo/tratamento farmacológico , Mieloma Múltiplo/patologia , Niacinamida/análogos & derivados , Compostos de Fenilureia/farmacologia , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Reguladoras de Apoptose/antagonistas & inibidores , Proteínas Reguladoras de Apoptose/genética , Western Blotting , Inibidores de Caspase/farmacologia , Caspases/química , Proliferação de Células/efeitos dos fármacos , Citometria de Fluxo , Humanos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Mieloma Múltiplo/metabolismo , Necrose , Niacinamida/farmacologia , Proteínas Proto-Oncogênicas/antagonistas & inibidores , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas c-bcl-2/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sorafenibe , Células Tumorais CultivadasRESUMO
Introducción: Los mucoceles son pseudotumores expansivos de las cavidades paranasales, cuyos síntomas están dados por su crecimiento expansivo. El tratamiento tradicional ha sido la extracción quirúrgica por vía externa, siendo desplazado en las últimas décadas por la resección endoscópica. Objetivo: Evaluar la experiencia en nuestro servicio sobre el diagnóstico y tratamiento de los mucoceles. Material y método: Revisión retrospectiva de pacientes tratados por mucocele en el Servicio de Otorrinolaringología, entre 2002y 2013. Se describen características clínico-demográficas, métodos diagnósticos, tratamiento y aparición de recidivas. Resultados: Se analizaron 45pacientes con seguimiento promedio de 24,93 meses. El 51,11% correspondió al sexo masculino. La edad promedio fue 49,68 años. El 46,6% fueron mucoceles fronto-etmoidales (6 frontales, 6 etmoidales y 9 fronto-etmoidales), el resto en seno maxilar, con un periodo de latencia de 7,33 meses. Los síntomas más frecuentes fueron oftalmológicos (55,5%) como proptosis y diplopía, mientras que el 51,1% de los pacientes tenía antecedentes de rinosinusitis crónica. El tratamiento fue mediante resección endoscópica en el 66,22%, por vía abierta 31,1% y manejo combinado en 6,66%. Hubo 8 casos recidivados, que representan el 17,7% de la serie. Discusión: Nuestra casuística resultó semejante a otras series publicadas. En los últimos años se ha preferido el abordaje endoscópico, sin embargo, el abordaje externo o combinado siguen siendo claves en el manejo de mucoceles extensos o para tratar recidivas. La aparición potencialmente tardía de recidivas requiere un tiempo de seguimiento largo para estos pacientes. Conclusiones: Los mucoceles representan un desafío diagnóstico y terapéutico y el manejo endoscópico parece una alternativa segura en el manejo de mucoceles que no comprometan órbita o tengan extensión craneal.
Introduction: Mucoceles are expansive pseudotumors of the paranasal sinuses and clinical presentations of symptoms are given by expansive growth. Traditional treatment has been open surgical extraction, being shifted in recent decades by endoscopic resection. Aim: To evaluate our experience in diagnosis and treatment of mucoceles. Material and methods: Retrospective chart review of patients treated for mucocele in the Department of Otolaryngology, between 2002 and 2013. Clinical and demographic characteristics, diagnostic methods, treatment and recurrence are described. Results: 45 patients with average follow-up of 24.93 months were analyzed. 51.11% were male. The mean age was 49.68years. 46.6% were fronto-ethmoidal mucoceles (6 in frontal sinus, 6 in ethmoidal sinus and 9 fronto-ethmoidal), the rest in the maxillary sinus, with a latency period of 7.33 months. Most frequent symptoms were ophtalmologic (55.5%) as proptosis and diplopia, while 51.1% of patients had a history of chronic rhinosinusitis. Treatment was by endoscopic resection in 66.22%, 31.1% by open surgery and 6.66% in combined approach. There were 8 recurrent cases, accounting for 17.7% of the series. Discussion: Our series was similar to other published earlier. In recent years, endoscopic approach is preferred, however, combined external approach or keys remain in handling large mucoceles or to treat recurrences. The late onset of recurrences potentially requires a long follow-up time for these patients. Conclusions: Mucoceles represent a diagnostic and therapeutic challenge and endoscopic management seems a safe alternative in the management of orbital involvement or cranial extension mucoceles.