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BACKGROUND: The purpose of this retrospective study was to evaluate treatment response, adverse events and survival rates of patients with intermediate stage HCC treated with superselective doxorubicin-loaded DC Bead transarterial chemoembolization (DEBDOX) under cone beam computed tomography (CBCT) control. PATIENTS AND METHODS: Between October 2010 and June 2012, 35 consecutive patients with intermediate stage HCC (32 male, 3 female; average age, 67.5 ± 7.8 years; 22 patients Child-Pugh class A, 8 class B, 5 without cirrhosis) were treated with DEBDOX TACE. Portal vein thrombosis was observed in 6 (17.1%) patients. DEBDOX TACE was performed by superselective catheterization of feeding vessels followed by embolization with 100-300 µm microspheres loaded with 50-100 mg of doxorubicin. In all cases, CBCT was used during chemoembolization. Tumor response rates were defined according to mRECIST criteria. RESULTS: Overall, 120 procedures were performed (mean, 3.2 per patients). We treated 97 lesions with an average diameter of 4.9 ± 1.9 cm. There were 32 minor and 2 (1.6%) major complications (one liver abscess and one cerebrovascular insult). After a mean follow-up of 27.7 ± 10.5 months, 94.3% of patients achieved an objective response to treatment (42.4% complete response and 57.6% partial response). Mean time to progression was 10.9 ± 5.3 months. Mean overall survival was 33.9 months (95% CI; 28.9 - 38.9 months), with 1- and 2- year survival of 97.1% and 65.7%, respectively. CONCLUSIONS: Superselective DEBDOX TACE performed under CBCT control is a safe and effective method with high rates of tumor response and overall survival.
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Clinical studies have indicated that the NV (nanovesicle) concentration in blood samples is a potential indicator of clinical status and can be used to follow the development of the disease. For 32 months, we monitored the effect of imatinib treatment on NV concentrations in blood samples from 12 patients with GIST (gastrointestinal stromal tumour). The NV concentration before the treatment increased with respect to control by a factor of 3.5 on average (range 2.6-9.2). The first week after initiation of the treatment, the NV concentration increased considerably, by a factor of 13 on average (range 5.9-21.2), whereas on average, after 1 month, it decreased to the level of the control and remained at that level for at least 1.5 years. Recent assessment (after 2.5 years) showed a somewhat increased NV concentration, by a factor of 2 on average (range 0.7-3.9). Low NV concentrations in blood samples during the treatment reflect a favourable effect of imatinib in these patients and no remission of the disease was hitherto observed.
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Antineoplásicos/uso terapêutico , Benzamidas/uso terapêutico , Tumores do Estroma Gastrointestinal/sangue , Tumores do Estroma Gastrointestinal/tratamento farmacológico , Piperazinas/uso terapêutico , Pirimidinas/uso terapêutico , Apoptose , Seguimentos , Tumores do Estroma Gastrointestinal/patologia , Humanos , Mesilato de Imatinib , Microscopia Eletrônica de VarreduraRESUMO
OBJECTIVES/HYPOTHESIS: Laryngopharyngeal reflux (LPR) has been proposed both as a trigger for recurrent respiratory papillomatosis (RRP) onset and as a factor favoring an aggressive clinical course. STUDY DESIGN: In this prospective study, 106 participants were recruited within a period of 24 months at the Department of Otorhinolaryngology and Cervicofacial Surgery, University Medical Centre Ljubljana. METHODS: This study compared a group of RRP patients (N = 36) with a group of LPR patients (N = 28) and a group of healthy participants (N = 42) based on Reflux Symptom Index (RSI), Reflux Finding Scores (RFS), and saliva analyses (pH, pepsin concentration, bile acid concentration, and pepsin enzymatic activity). RESULTS: The RRP group compared to the LPR group showed a statistically significant difference only in RSI and RFS scores, while the RRP group compared to healthy controls showed significantly higher values in all tested parameters (RSI score, RFS, saliva pH, pepsin concentration, bile acids concentration, pepsin enzymatic activity). CONCLUSIONS: LPR is common in RRP patients and significantly more prevalent compared to healthy controls. Our results show that saliva analyses are a better office-based tool than RSI questionnaires and RFS scores for diagnosing LPR in RRP patients. LEVEL OF EVIDENCE: 3 Laryngoscope, 132:619-625, 2022.
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Refluxo Laringofaríngeo/complicações , Infecções por Papillomavirus/etiologia , Infecções Respiratórias/etiologia , Adulto , Fatores Etários , Estudos de Casos e Controles , Feminino , Humanos , Concentração de Íons de Hidrogênio , Refluxo Laringofaríngeo/diagnóstico , Masculino , Infecções por Papillomavirus/epidemiologia , Infecções por Papillomavirus/patologia , Estudos Prospectivos , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/patologia , Fatores de Risco , Saliva/químicaRESUMO
BACKGROUND: 13C-mixed triglyceride breath test (13C-MTGT) is a non-invasive test for the detection of moderate and severe pancreatic exocrine insufficiency (PEI), but it requires prolonged breath sampling. The aim of this study was to determine the diagnostic power of abbreviated 13C-MTGT in detecting PEI in patients after subtotal and total gastrectomy performed due to gastric cancer. SUBJECTS AND METHODS: This cross-sectional observational study included 3 groups of subjects; healthy controls, patients with subtotal and patients with total gastrectomy. Demographic and clinical data of patients were collected. Stool samples to determine faecal elastase (Fe-1) and chymotrypsin were collected and measured by ELISA. All subjects performed 5-hour 13C-MTGT breath test. The concentration and relative content of 13C in exhaled air was measured by isotope ratio mass spectrometer (IRMS). PEI was confirmed as values of 13C-exhalation < 26.8% after 5 hours. RESULTS: Overall, 65 participants were included into analysis, 22 having PEI (n = 11 after subtotal and n = 11 after total gastrectomy, both performed for gastric cancer). 13C-MTGT breath test showed difference in percent of exhaled 13C between PEI and non-PEI patients already after 60 minutes (p = 0.034). Receiver operating characteristic (ROC) curve analysis showed that cut-off value of 13.74% after 150 minutes is showing equivalent diagnostic power to the longer test with sensitivity and specificity both above 90% for the exclusion of PEI in patients after subtotal and/or total gastrectomy. CONCLUSIONS: In this study abbreviated 13C-MTGT test could be shortened from 5 to 2.5 hours without decrease in its diagnostic accuracy for detection of PEI in patients with subtotal or total gastrectomy performed for gastric cancer. This allows significant time savings in the diagnostics of PEI in this subgroup of patients.
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Insuficiência Pancreática Exócrina , Neoplasias Gástricas , Testes Respiratórios , Estudos Transversais , Insuficiência Pancreática Exócrina/diagnóstico , Insuficiência Pancreática Exócrina/etiologia , Gastrectomia , Humanos , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/cirurgia , TriglicerídeosRESUMO
BACKGROUND: Drug-eluting microsphere transarterial chemoembolization (DEM-TACE) is the standard of care in patients with intermediate-stage hepatocellular carcinoma and ensures targeted and controlled cytotoxic and ischemic effects. Proper patient selection and optimized treatment techniques are associated with longer median survival. The aim of this single-institution retrospective study was to evaluate safety and efficacy of DEM-TACE under cone beam computed tomography (CBCT) control in patients with early and intermediate stage hepatocellular carcinoma. PATIENTS AND METHODS: A total of 144 patients (mean age 67.9 ± 8.0 years, 127 males and 17 females) between February 2010 and December 2018 were studied. Microparticles of different dimensions according to two manufacturers (diameter of 70-150 µm, 100-300 µm or 300-500 µm and 40-µm, 75-µm or 100-µm) were used and loaded with 50-150 mg of doxorubicin. The objective tumour response according to the modified Response Evaluation Criteria in Solid Tumours (mRECIST), the time to progression, adverse events and overall survival were (OS) evaluated. RESULTS: In total, 452 procedures were performed (median, 3 per patient). Four (0.9% of all procedures) major complications were noted. Postembolization syndrome occurred after 35% of procedures. At the first imaging follow-up 2-3 months after first treatment, 91% of patients achieved an objective response. The median time to progression was 10.2 months (95% CI: 8.3-12.1 months). OS rates at 1, 2, 3, 4, and 5 years were 85%, 53%, 33%, 20% and 14%, respectively. The median survival time was 25.8 months (95% CI: 22.1-29.5 months). CONCLUSIONS: DEM-TACE under CBCT control in patients with early and intermediate stage hepatocellular carcinoma is a safe and effective method of treatment with high objective tumour response and survival rates.
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Carcinoma Hepatocelular , Quimioembolização Terapêutica , Neoplasias Hepáticas , Idoso , Antibióticos Antineoplásicos , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/terapia , Quimioembolização Terapêutica/métodos , Tomografia Computadorizada de Feixe Cônico , Feminino , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/terapia , Masculino , Microesferas , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
BACKGROUND: Microvesicles isolated from body fluids are membrane - enclosed fragments of cell interior which carry information on the status of the organism. It is yet unclear how metabolism affects the number and composition of microvesicles in isolates from the peripheral blood. AIM: To study the post - prandial effect on microvesicles in isolates from the peripheral blood of 21 healthy donors, in relation to blood cholesterol and blood glucose concentrations. RESULTS: The average number of microvesicles in the isolates increased 5 hours post - prandially by 52%; the increase was statistically significant (p = 0.01) with the power P = 0.68, while the average total blood cholesterol concentration, average low density lipoprotein cholesterol concentration (LDL-C) and average high density lipoprotein cholesterol concentration (HDL-C) all remained within 2% of their fasting values. We found an 11% increase in triglycerides (p = 0.12) and a 6% decrease in blood glucose (p < 0.01, P = 0.74). The post - prandial number of microvesicles negatively correlated with the post - fasting total cholesterol concentration (r = - 0.46, p = 0.035) while the difference in the number of microvesicles in the isolates between post - prandial and post - fasting states negatively correlated with the respective difference in blood glucose concentration (r = - 0.39, p = 0.05). CONCLUSIONS: In a population of healthy human subjects the number of microvesicles in isolates from peripheral blood increased in the post - prandial state. The increase in the number of microvesicles was affected by the fasting concentration of cholesterol and correlated with the decrease in blood glucose.
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Glicemia/metabolismo , Colesterol/sangue , Endossomos/metabolismo , Adulto , Comunicação Celular , HDL-Colesterol/sangue , LDL-Colesterol/sangue , Jejum/sangue , Feminino , Citometria de Fluxo , Humanos , Masculino , Pessoa de Meia-Idade , Período Pós-Prandial , Triglicerídeos/sangueRESUMO
Microvesicles are sub-micron structures shed from the cell membrane in a final step of the budding process. After being released into the microenvironment they are free to move and carry signaling molecules to distant cells, thereby they represent a communication system within the body. Since all cells shed microvesicles, it can be expected that they will be found in different body fluids. The potential diagnostic value of microvesicles has been suggested, however, a standardized protocol for isolation has not yet been agreed upon. It is unclear what is the content of the isolates and whether the isolated microvesicles were present in vivo or-have they been created within the isolation procedure. To present evidence in this direction, in this work we focus on the visualization of the material obtained by the microvesicle isolation procedure. We present scanning electronic microscope images of microvesicles isolated from blood, ascites, pleural fluid, cerebrospinal fluid, postoperative drainage fluid and chyloid fluid acquired from human and animal patients. Vesicular structures sized from 1microm downto 50nm are present in isolates of all considered body fluids, however, the populations differ in size and shape reflecting also the composition of the corresponding sediments. Isolates of microvesicles contain numerous cells which indicates that methods of isolation and determination of the number of microvesicles in the peripheral blood are to be elaborated and improved.
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Sangue , Líquidos Corporais , Micropartículas Derivadas de Células/ultraestrutura , Microscopia Eletrônica de Varredura , Adenocarcinoma/complicações , Adenocarcinoma/veterinária , Idoso , Animais , Ascite/etiologia , Ascite/patologia , Carcinoma/sangue , Carcinoma/complicações , Doenças do Gato/patologia , Gatos , Quilotórax/patologia , Quilotórax/veterinária , Neoplasias do Colo/complicações , Feminino , Cavalos/sangue , Humanos , Neoplasias Pulmonares/complicações , Neoplasias Pulmonares/veterinária , Neoplasias Pancreáticas/sangue , Tamanho da Partícula , Peritonite/complicações , Derrame Pleural Maligno/etiologia , Derrame Pleural Maligno/patologia , Hemorragia Subaracnóidea/líquido cefalorraquidianoRESUMO
Background Sorafenib is an oral multi-kinase inhibitor used for the treatment of hepatocellular carcinoma. Its efficacy in randomised controlled trials was demonstrated in patients with well-preserved liver function and good functional status. In the real-world setting, treatment is often offered to patients outside these criteria. We therefore performed a single-centre real-world cohort study on the efficacy of sorafenib in patients with hepatocellular carcinoma. Patients and methods We identified all patients with hepatocellular carcinoma initiating treatment with sorafenib between January 2015 and January 2018. The primary endpoint was overall survival (OS) since starting sorafenib. Clinical and demographic variables associated with survival were studied. Results The median OS was 13.4 months (95% CI 8.2-18.6). Multivariable Cox's regression identified worse ECOG performance status (HR 2.21; 95% CI 1.56-3.16; P < 0.0001), Child-Pugh class C (HR 52.4; 95% CI 3.20-859; P = 0.005) and absence of prior locoregional treatment (HR 2.30; 95% CI 1.37-3.86; P = 0.002) to be associated with increased mortality. Conclusions Careful selection of patients for treatment with sorafenib is of paramount importance to optimize outcomes.
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Antineoplásicos/uso terapêutico , Carcinoma Hepatocelular/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Inibidores de Proteínas Quinases/uso terapêutico , Sorafenibe/uso terapêutico , Idoso , Carcinoma Hepatocelular/mortalidade , Estudos de Coortes , Feminino , Humanos , Neoplasias Hepáticas/mortalidade , Masculino , Pessoa de Meia-Idade , Seleção de PacientesRESUMO
The aim of this clinical study was to investigate the effectiveness and long-term safety of electrochemotherapy as an emerging treatment for HCC in patients not suitable for other treatment options. A prospective phase II clinical study was conducted in patients with primary HCC who were not suitable for other treatment options according to the Barcelona Clinic Liver Cancer classification. A total of 24 patients with 32 tumors were treated by electrochemotherapy. The procedure was effective, feasible, and safe with some procedure-related side effects. The responses of the 32 treated nodules were: 84.4% complete response (CR), 12.5% partial response (PR), and 3.1% stable disease (SD). The treatment was equally effective for nodules located centrally and peripherally. Electrochemotherapy provided a durable response with local tumor control over 50 months of observation in 78.0% of nodules. The patient responses were: 79.2% CR and 16.6% PR. The median progression-free survival was 12 months (range 2.7-50), and the overall survival over 5 years of observation was 72.0%. This prospective phase II clinical study showed that electrochemotherapy was an effective, feasible, and safe option for treating HCC in patients not suitable for other treatment options.
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Background Electrochemotherapy is an effective treatment of colorectal liver metastases and hepatocellular carcinoma (HCC) during open surgery. The minimally invasive percutaneous approach of electrochemotherapy has already been performed but not on HCC. The aim of this study was to demonstrate the feasibility, safety and effectiveness of electrochemotherapy with percutaneous approach on HCC. Patient and methods The patient had undergone the transarterial chemoembolization and microwave ablation of multifocal HCC in segments III, V and VI. In follow-up a new lesion was identified in segment III, and recognized by multidisciplinary team to be suitable for minimally invasive percutaneous electrochemotherapy. The treatment was performed with long needle electrodes inserted by the aid of image guidance. Results The insertion of electrodes was feasible, and the treatment proved safe and effective, as demonstrated by control magnetic resonance imaging. Conclusions Minimally invasive, image guided percutaneous electrochemotherapy is feasible, safe and effective in treatment of HCC.
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Bleomicina/uso terapêutico , Carcinoma Hepatocelular/tratamento farmacológico , Eletroquimioterapia/métodos , Neoplasias Hepáticas/tratamento farmacológico , Técnicas de Ablação , Idoso , Angiografia , Carcinoma Hepatocelular/diagnóstico por imagem , Terapia Combinada , Tomografia Computadorizada de Feixe Cônico , Humanos , Neoplasias Hepáticas/diagnóstico por imagem , Masculino , Micro-Ondas/uso terapêutico , Radiografia IntervencionistaRESUMO
Heparins (unfractionated and low molecular weight (LMWH) heparins) primarily used as anticoagulants, were found to be effective also in slowing down the development of some types of cancer. On the other hand, the number of microvesicles in the peripheral blood originating from the budding of cell membranes (mostly platelets) is increased in hypercoagulabile states as well as in cancer, indicating a possible common underlying mechanism. It was hypothesized that by mediating an attractive interaction between phospholipid membranes heparin suppresses microvesiculation and thereby acts as an anticoagulant and anti-tumor agent. In this work, the effect of LMWH nadroparin on phospholipid membranes was tested in vitro in a system of giant phospholipid vesicles (GPVs) created by electroformation and observed under the phase contrast microscope. Plasma of different blood donors containing different concentrations of nadroparin was added to the suspension of GPVs to induce adhesion between GPVs. The attractive interaction between membranes was assessed by measuring the average effective angle of contact between the adhered GPVs. It was found in healthy donors, in a donor with gastrointestinal cancer and in a donor with rheumatoid arthritis that adding therapeutic doses of nadroparin to the plasma samples enhanced adhesion of phospholipid membranes in a dose and time-dependent manner while nadroparin alone had no effect within the therapeutic concentration range. The results are in favor of the hypothesis that suppression of microvesiculation underlies both, the anticoagulant and the anti-tumor progression effect of heparin.
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Anticoagulantes/farmacologia , Micropartículas Derivadas de Células/efeitos dos fármacos , Nadroparina/farmacologia , Trombofilia/tratamento farmacológico , Adulto , Idoso , Anticoagulantes/uso terapêutico , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Artrite Reumatoide/sangue , Artrite Reumatoide/tratamento farmacológico , Cardiolipinas/química , Feminino , Neoplasias Gastrointestinais/sangue , Neoplasias Gastrointestinais/tratamento farmacológico , Humanos , Lipossomos/química , Masculino , Lipídeos de Membrana/química , Microscopia de Contraste de Fase , Pessoa de Meia-Idade , Nadroparina/uso terapêutico , Fosfatidilcolinas/química , Fosfolipídeos/química , Plasma , Soluções , Sacarose/farmacologia , Trombofilia/sangue , Trombofilia/etiologiaRESUMO
It was recently shown that the plasma protein-mediated attractive interaction between phospholipid membranes could in the budding process cause adhesion of the bud to the mother membrane [J. Urbanija, N. Tomsic, M. Lokar, A. Ambrozic, S. Cucnik, M. Kanduser, B. Rozman, A. Iglic, V. Kralj-Iglic, Coalescence of phospholipid membranes as a possible origin of anticoagulant effect of serum proteins, Chem. Phys. Lipids 150 (2007) 49-57]. Since in the in vivo conditions the budding of cell membranes leads to the release of microvesicles into the circulation, a hypothesis was put forward that the ability of plasma to cause adhesion between membranes supresses the microvesiculation process. In the present work, this hypothesis was tested in a population of 19 patients with gastrointestinal diseases. The number of microvesicles in peripheral blood of patients was determined by flow cytometry while the ability of plasma to cause adhesion between membranes was determined by adding patient's plasma to the suspension of giant phospholipid vesicles created by electroformation method, and measuring the average effective angle of contact between the adhered vesicles. Statistically significant negative correlations between the number of microvesicles and the average effective angle of contact (Pearson coefficient -0.50, p=0.031) and between the number of microvesicles per number of platelets and the average effective angle of contact (Pearson coefficient -0.64, p=0.003) were found, which is in favor of the above hypothesis. Patients with gastrointestinal cancer had larger number of microvesicles (difference 140%, statistical significance 0.033) and smaller average effective angle of contact (difference 20%, statistical significance 0.013) compared to patients with other gastrointestinal diseases.
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Plaquetas/fisiologia , Membrana Celular/fisiologia , Vesículas Citoplasmáticas/fisiologia , Gastroenteropatias/fisiopatologia , Plasma/fisiologia , Adesividade , Idoso , Idoso de 80 Anos ou mais , Feminino , Gastroenteropatias/sangue , Gastroenteropatias/patologia , Humanos , Lipossomos/metabolismo , Masculino , Pessoa de Meia-Idade , Fosfolipídeos/fisiologia , Contagem de PlaquetasRESUMO
During harvesting of nanovesicles (NVs) from blood, blood cells and other particles in blood are exposed to mechanical forces which may cause activation of platelets, changes of membrane properties, cell deformation and shedding of membrane fragments. We report on the effect of shear forces imposed upon blood samples during the harvesting process, on the concentration of membrane nanovesicles in isolates from blood. Mathematical models of blood flow through the needle during sampling with vacuumtubes and with free flow were constructed, starting from the Navier-Stokes formalism. Blood was modeled as a Newtonian fluid. Work of the shear stress was calculated. In experiments, nanovesicles were isolated by repeated centrifugation (up to 17,570×g) and washing, and counted by flow cytometry. It was found that the concentration of nanovesicles in the isolates positively corresponded with the work by the shear forces in the flow of the sample through the needle. We have enhanced the effect of the shear forces by shaking the samples prior to isolation with glass beads. Imaging of isolates by scanning electron microscopy revealed closed globular structures of a similar size and shape as those obtained from unshaken plasma by repetitive centrifugation and washing. Furthermore, the sizes and shapes of NVs obtained by shaking erythrocytes corresponded to those isolated from shaken platelet-rich plasma and from unshaken platelet rich plasma, and not to those induced in erythrocytes by exogenously added amphiphiles. These results are in favor of the hypothesis that a significant pool of nanovesicles in blood isolates is created during their harvesting. The identity, shape, size and composition of NVs in isolates strongly depend on the technology of their harvesting.
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Eritrócitos/citologia , Vesículas Extracelulares , Nanoestruturas , Agulhas , Plasma Rico em Plaquetas/citologia , Adulto , Vesículas Extracelulares/ultraestrutura , Feminino , Citometria de Fluxo , Humanos , Masculino , Microscopia Eletrônica de Varredura , Pessoa de Meia-Idade , Nanoestruturas/ultraestrutura , Estresse Mecânico , Adulto JovemRESUMO
Gastroesophageal reflux (GER) can cause serious voice problems and laryngopharyngeal disorders influencing the patient's quality of life. Forty-three patients with suspected laryngopharyngeal reflux (LPR) were included into a prospective study. The diagnosis was made on the basis of the patient's history, the videoendolaryngoscopy, the oesophago-gastroscopy and the biopsy of the oesophageal mucosa. All the LPR patients were treated with esomeprasol for eight weeks. An acoustic analysis of the vowel /a/ samples was performed in the LPR group before and after the treatment. Thirty-six patients with vocal fold polyps served as the control group for a subjective estimation of the voice problems. All the patients from both groups subjectively evaluated their voice problems using the Voice Handicap Index (VHI) questionnaire. The results of VHI showed that the severity of the voice problems of the patients with LPR could be compared to that experienced by the patients with vocal fold polyps. Videoendolaryngoscopy and history proved LPR in all 43 patients. Oesophago-gastroscopy combined with the histopathological examination of the oesophageal biopsy specimens detected signs of possible GER in 38 patients (88%). The results of the videoendolaryngoscopy combined with a subjective and objective voice assessment, performed before and after treatment with a proton-pump inhibitor, showed a significant improvement in most of the studied parameters by the end of the therapy. In the diagnostics of LPR, the patient's history and videoendolaryngoscopy demonstrated to be superior to oesophago-gastroscopy. Videoendolaryngoscopic assessment of the laryngeal mucosa, and oesophago-gastroscopy supplemented with a biopsy of the oesophageal mucosa, showed to be a convenient diagnostic method when GER and LPR were suspected. Esomeprasol proved to be very effective in the treatment of LPR. LPR should not be overlooked in the treatment of dysphonic patients.
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Refluxo Gastroesofágico/complicações , Refluxo Gastroesofágico/diagnóstico , Distúrbios da Voz/etiologia , Adolescente , Adulto , Idoso , Estudos de Casos e Controles , Inibidores Enzimáticos/uso terapêutico , Esofagoscopia , Feminino , Refluxo Gastroesofágico/tratamento farmacológico , Gastroscopia , Humanos , Doenças da Laringe/complicações , Laringoscopia , Masculino , Pessoa de Meia-Idade , Omeprazol/uso terapêutico , Pólipos/complicações , Estudos Prospectivos , Inibidores da Bomba de Prótons , Qualidade de Vida , Índice de Gravidade de Doença , Inquéritos e Questionários , Distúrbios da Voz/psicologiaRESUMO
BACKGROUND: Shedding of nanoparticles from the cell membrane is a common process in all cells. These nanoparticles are present in body fluids and can be harvested by isolation. To collect circulating nanoparticles from blood, a standard procedure consisting of repeated centrifugation and washing is applied to the blood samples. Nanoparticles can also be shed from blood cells during the isolation process, so it is unclear whether nanoparticles found in the isolated material are present in blood at sampling or if are they created from the blood cells during the isolation process. We addressed this question by determination of the morphology and identity of nanoparticles harvested from blood. METHODS: The isolates were visualized by scanning electron microscopy, analyzed by flow cytometry, and nanoparticle shapes were determined theoretically. RESULTS: The average size of nanoparticles was about 300 nm, and numerous residual blood cells were found in the isolates. The shapes of nanoparticles corresponded to the theoretical shapes obtained by minimization of the membrane free energy, indicating that these nanoparticles can be identified as vesicles. The concentration and size of nanoparticles in blood isolates was sensitive to the temperature during isolation. We demonstrated that at lower temperatures, the nanoparticle concentration was higher, while the nanoparticles were on average smaller. CONCLUSION: These results indicate that a large pool of nanoparticles is produced after blood sampling. The shapes of deformed blood cells found in the isolates indicate how fragmentation of blood cells may take place. The results show that the contents of isolates reflect the properties of blood cells and their interaction with the surrounding solution (rather than representing only nanoparticles present in blood at sampling) which differ in different diseases and may therefore present a relevant clinical parameter.