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BACKGROUND: Personality traits as alexithymia and type D personality may impair health related quality of life (HRQoL) in patients with irritable bowel syndrome (IBS) and inflammatory bowel disease (IBD). Aim of this study was to evaluate personality traits in patients with IBS and IBD and their impact on HRQoL. MATERIALS AND METHODS: The subjects (40 patients with IBS, 40 with IBD and 40 health control subjects) completed SF-36 questionnaire, TAS-20 and DS14 scale. RESULTS: Patients with IBS and IBD had higher results on TAS-20 and DS14 scale when compared with healthy controls. Also IBS patients had higher scores than IBD patients. Higher scores on TAS-20 and DS14 scales in IBS and IBD patients correlate with lower HRQoL. HRQoL was poorer in IBS and IBD patients than in healthy control subjects. CONCLUSIONS: Alexithymia and type D personality in IBS and IBD patients are associated with lower HRQoL and psychological interventions should be considered.
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Enfermedades Inflamatorias del Intestino , Síndrome del Colon Irritable , Humanos , Síndrome del Colon Irritable/complicaciones , Calidad de Vida/psicología , Enfermedades Inflamatorias del Intestino/complicaciones , Enfermedades Inflamatorias del Intestino/psicología , Encuestas y Cuestionarios , PersonalidadRESUMEN
Recent studies have established a concept of tumour necrosis factor-α (TNF-α)/Fas signalling crosstalk, highlighting TNF-α as a critical cytokine in sensitizing hepatocytes to death induced by Fas activation. However, in the exact inflammatory response, besides TNF-α, many other mediators, that might modulate apoptotic response differentially, are released. To resolve the issue, we studied the effects of lipopolysaccharide (LPS), one of the crucial inductors of inflammation in the liver, on apoptotic outcome. We show that LPS-induced inflammation diminishes the sensitivity of hepatocytes to Fas stimulus in vivo at caspase-8 level. Analysis of molecular mechanisms revealed an increased expression of various pro-inflammatory cytokines in non-parenchymal liver cells and hepatocyte-specific increase in Bcl-xL, associated with signal transducer and activator of transcription 3 (Stat3) phosphorylation. Pre-treatment with ruxolitinib, a selective Janus kinase (JAK) 1/2 inhibitor, prevented the LPS-induced Stat3 phosphorylation and restored the sensitivity of hepatocytes to Fas-mediated apoptosis. Furthermore, ruxolitinib pre-treatment diminished the LPS-induced Bcl-xL up-regulation without an inhibitory effect on LPS-induced expression of pro-inflammatory cytokines. In summary, although the reports are showing that the effects of isolated pro-inflammatory mediators, such as TNF-α or neutrophils, are pro-apoptotic, the overall effect of inflammatory milieu on hepatocytes in vivo is Stat3-dependent desensitization to Fas-mediated apoptosis.
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Inflamación/tratamiento farmacológico , Pirazoles/farmacología , Factor de Transcripción STAT3/genética , Factor de Necrosis Tumoral alfa/genética , Receptor fas/genética , Animales , Apoptosis/efectos de los fármacos , Caspasa 8/genética , Hepatocitos/efectos de los fármacos , Hepatocitos/patología , Humanos , Inflamación/inducido químicamente , Inflamación/genética , Inflamación/patología , Lipopolisacáridos/toxicidad , Hígado/efectos de los fármacos , Hígado/patología , Ratones , Nitrilos , Pirimidinas , Factor de Transcripción STAT3/antagonistas & inhibidores , Transducción de Señal/efectos de los fármacos , Proteína bcl-X/genéticaRESUMEN
INTRODUCTION: During the plateletpheresis procedure the number of thrombocytes in the donor's blood significantly decreases, and the levels of the hematocrit (HCT), hemoglobin (Hgb), and leukocyte (WBC) diminish as well. Influence of the cell separator is one of the factors that affects the levels of HCT, Hgb and WBC. In this study, the goal was to determine the value difference of HCT, Hgb, WBC, and platelets after the platelet pheresis process between performance on Fenwal AMICUS and on Fresenius Com Tec. DONORS AND METHODS: The criteria for participation: male in the age range of 25-45. We have formed two groups: for both groups - 180 separations were performed on 60 participants were the values of hematocrits, concentration of hemoglobin and number of leukocytes were established before and after separation using the double-needle continuous flow cell separation (DN-CFCS) on two different devices, Fenwal AMICUS device and the Fresenius Com Tec. device. To confirm the statistical differences we have used Student t-test for independent or dependent samples, as well as Mann-Whitney U test as non-parametric alternative. To compare differences between the values of four parameters (P1-P2) from two groups (using two devices - Fenwal AMICUS and Fresenius Com Tec) The possibility of errors were accepted for α<0.05, and the difference between groups as statistical relevant were accepted for p<0.05. RESULTS: Statistically significant lower values were noted for all researched parameters after separation on both devices. The statistically significant average values for Hct, Hgb and WBC obtained between two devices, were less than 0.05 (p=0.05). For the platelets (Plt) there was no statistical significant difference (p>0.05 - α=0.05), between average level obtained using either Fenwal AMICUS or Frazenius Com Tec. CONCLUSION: The type of cell separator had the influence on the decrease value of the observed parameters.
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Células Sanguíneas/citología , Donantes de Sangre , Separación Celular/instrumentación , Separación Celular/métodos , Adulto , Plaquetas/citología , Humanos , Leucocitos/citología , Masculino , Persona de Mediana Edad , PlaquetoferesisRESUMEN
AIM: To assess the prognostic value of lactate level for mortality in patients with pulmonary embolism (PE) and Pulmonary Embolism Severity Index (PESI) I-III and its independence of gas-analysis parameters and acid-base status. METHODS: This prospective observational study was conducted at the University Clinical Hospital Mostar from 2013 to 2017. On the first day after PE diagnosis, 1.5 mL of arterial blood was collected from 103 patients with PE. Partial pressure of oxygen in arterial blood, partial pressure of carbon dioxide in arterial blood, blood pH value, concentration of bicarbonates in arterial blood (HCO3-), base deficit, and oxygen saturation were analyzed. Lactate levels were assessed using blood samples taken from the cubital vein. Logistic regression analysis was used to assess the predictive value of gas-analysis variables, lactate level, PESI score, age, and sex for in-hospital death due to PE. RESULTS: The mortality in the group of PE patients was 19.1% (18 of 103 patients). Lactate level was an independent predictor of mortality (P=0.002, odds ratio 0.06). HCO3- was also found to be a significant predictor (P=0.022, odds ratio 2.4). Lactates were independent of other variables. Other gas-analysis parameters were not significant predictors of mortality. CONCLUSION: In PE patients at low-intermediate risk of mortality (PESI I-III), lactate level was associated with a short-term mortality, independently of other gas-analytic parameters. Oxford Centre for Evidence-based Medicine level of evidence: 2.
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Equilibrio Ácido-Base/fisiología , Lactatos/sangre , Embolia Pulmonar/sangre , Embolia Pulmonar/mortalidad , Anciano , Análisis de los Gases de la Sangre , Dióxido de Carbono/sangre , Femenino , Humanos , Concentración de Iones de Hidrógeno , Masculino , Persona de Mediana Edad , Oportunidad Relativa , Oxígeno/sangre , Valor Predictivo de las Pruebas , Pronóstico , Estudios Prospectivos , Índice de Severidad de la EnfermedadRESUMEN
INTRODUCTION: During the plateletpheresis procedure the number of trombocites in the donor's blood significantly decreases, and the levels of the other components of blood as hematocrit, hemoglobin, and leukocyte diminish as well. Influence of the type of procedure DN-CFCS and SN-ICFS it is one of the factors that affects the decrease of the levels of HCT, Hgb and WBC. In this study, our goal was to see the difference in the value of HCT, Hgb, WBC, and platelets after the plateletfphresis process between DN-CFCS and SN-IFCS on the same cell separator - Fenval AMICUS. DONORS AND METHODS: The criteria for participation: men between age of 25-45. Two groups were formed. Group I 112 separation done with the method SN-ICFS and Group II 180 separation done with the method DN-CFCS. STATISTICAL ANALYSIS: To confirm the statistical difference we used Student t-test for independent or dependent samples, as well as Mann-Whitney U test as non parametric alternative. The possibility of errors were accepted for α<0.05, and the difference between groups were accepted as statistical relevant for p<0.05. RESULTS: Statistically significant lower values were observed of all researched parameters after separation for the donors on the equipement Amicus DN, and for donors on Amicus SN. A significant higher value of HCT before procedure was found in the AM DN group, in the researches of the other variables there were no significant differences. The resultst for the comparison of variables after procedure procedure for DN and SN procedure. A significant higher value of HCT and a significant higher level of Hgb, as well as a significant lower level of WBV after procedure in the AM DN group, while for the levels of PLT there were no significant differences. CONCLUSION: On the decrease of the value of the observed parameters the type of procedure has an influence that means DN-CFCS or SN-IFCS, continuous or discontinuous flow.
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Donantes de Sangre , Plaquetas/citología , Plaquetoferesis/métodos , HumanosRESUMEN
Over the years, an electrocardiogram had become the basic tool to study the heart physiology and pathophysiology. Many authors gave a substantial contribution in understanding the electrophysiological basis for numerous electrocardiographic changes. Some of them were named after authors themselves, or others used the names of scientists who first discovered or explained the nature of a particular electrocardiographic finding. In this article, electrocardiographic phenomena and eponyms used in today's electrocardiography are described.
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Arritmias Cardíacas/clasificación , Arritmias Cardíacas/historia , Electrocardiografía/clasificación , Electrocardiografía/historia , Terminología como Asunto , Arritmias Cardíacas/diagnóstico , Historia del Siglo XIX , Historia del Siglo XX , Historia del Siglo XXI , Humanos , InternacionalidadRESUMEN
AIM: The main goal of this study was to compare the biochemical and histopathological findings in patients with sustained virological response (SVR) before and two years after the therapy with pegylated interferon α-2a and ribavirin in chronic hepatitis C. SUBJECTS AND METHODS: The study was conducted at the Department of Internal Medicine and the Clinic for Infectious Diseases of the Clinical Hospital Mostar. The study included 48 patients whose treatment for chronic hepatitis C with pegylated interferon α-2a and ribavirin was finished two years prior to the achieved SVR at the end of the treatment. The main criterion for inclusion was a negative result of HCV RNA, determined by the RealTime HCV assay. After taking a history, physical examination, laboratory tests: AST, ALT, GGT, a liver biopsy were performed with the help of the ultrasound. The assessment of necroinflamatory score was determined by histologic activity index (HAI) score, and the stage of fibrosis according to Knodell's numerical score. RESULTS: The values of AST and ALT levels were statistically significantly decreased after the successful treatment (p<0.001), as well as the value of HAI score (p=0.001) and the stage of fibrosis (p=0.010), in contrast to GGT (p=0.054). For the components of HAI score like focal necrosis (0.001) and portal inflammation (0.042) the result showed that they were significantly higher before the therapy, which was not true for the piecemeal (p=0.054) and confluated necrosis (p=0.078). The improvement of HAI score after therapy was found in 36 patients (75.0%), and 27 patients (56.2%) showed an improvement in the degree of fibrosis with the most common improvement of 1 degree (85.7%). One third of patients (31.3%) had the same result in the degree of fibrosis before and after the therapy. Before the treatment, a positive correlation was observed between ALT (p=0.039) and AST (p=0.04) with HAI, AST and the stage of fibrosis (p=0.04). In contrast, after the treatment the only correlation was observed between AST and the stage of fibrosis (p=0.042). CONCLUSION: Virological and biochemical responses in patients with SVR may not reflect the histopathological effects of the treatment and therefore these patients should be monitored for the possible development of the liver cirrhosis and hepatocellular carcinoma.
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OBJECTIVES: The aim of this study was to determine the disturbances in the concentration of parathyroid hormone (PTH) and 25-hydroxyvitamin D (vitamin D) in patients with stable chronic obstructive pulmonary disease (COPD) and its correlation with airflow obstruction. MATERIALS AND METHODS: A prospective study included 200 patients with a confirmed diagnosis of COPD in the Department of Lung Diseases and Tuberculosis and Pulmonology Polyclinic of University Clinical Hospital Mostar in the period of three years, between May 2021 and May 2024. Inclusion criteria were a stable phase of COPD, hemodynamically stable patients older than 40 years, forced vital capacities in the first second (FEV1)/forced vital capacities (FVC) <0.7, and patients with PTH, vitamin D, calcium, and phosphate measurements. Exclusion criteria were acute exacerbation of COPD in the last month; current treatment with nutritional supplements, vitamins, and statins; lack of availability of lung function data; use of systemic corticosteroids in the previous three months; chronic renal insufficiency, respiratory diseases other than COPD (asthma, pneumonia, tuberculosis, and bronchiectasis), and other diseases (cancer and parathyroid disease). Medical records about demographic data (age and gender), pulmonary function test (FVC, FEV1, FEV1%FVC, mean expiratory flow (MEF)50), body mass index (BMI), COPD assessment test (CAT), Modified Medical Research Council (mMRC) Dyspnea Scale, and serum PTH, vitamin D, calcium, and phosphate levels were obtained. RESULTS: Patients with higher COPD stage had lower spirometry values, most significantly MEF50. The higher the COPD group (Global Initiative for Chronic Obstructive Lung Disease (GOLD) D), the lower vitamin D ââand the higher PTH levels were. Calcium and phosphate values ââwere the same for all groups. Vitamin D and PTH levels significantly ââcorrelated with MEF50 values. The lower MEF50 level, the higher PTH levels, ââand lower vitamin D levels were found (P<0.05). CONCLUSION: Our study showed that the patients in the higher COPD group have lower vitamin D levels ââand higher PTH levels, indicating that they developed secondary hyperparathyroidism. The levels of vitamin D and PTH correlated the most with MEF50 values while other spirometry parameters did not significantly correlate with vitamin D and PTH levels.
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OBJECTIVES: The aim of this study was to determine the short-term consequences of coronavirus disease 2019 (COVID-19) infection on pulmonary diffusion in patients with severe (but not critical) and moderately severe COVID-19 pneumonia during three months after COVID-19 infection. METHODS: A prospective study included 81 patients with an RT-PCR-test confirmed diagnosis of COVID-19 infection treated in the COVID Department of Lung Diseases of University Clinical Hospital Mostar. Inclusion criteria were ≥18-year-old patients, COVID-19 infection confirmed using real-time RT-PCR, radiologically confirmed bilateral COVID-19 pneumonia, and diffusion capacity of the lungs for carbon monoxide (DLCO) one and three months after COVID-19 infection. The pulmonary function was tested using the MasterScreen Body Jaeger (Jaeger Corporation, Omaha, USA) and MasterScreen PFT Jaeger (Jaeger Corporation, Omaha, USA) according to American Thoracic Society guidelines one and three months after COVID-19 infection. RESULTS: Forced vital capacity significantly increased three months after COVID-19 infection compared to the first-month control (p<0.0005). Also, a statistically significant increase in the FEV1 value (p<0.0005), FEV1%FVC ratio (p<0.005), DLCO/SB (p<0.0005), DLCO/VA value (p<0.0005), and total lung capacity (TLC) (p<0.0005) was observed in all patients. CONCLUSION: Our study showed that recovery of DLCO/VA and spirometry parameters was complete after three months, while DLCO/SB was below normal values even after three months. Therefore, one month after the COVID-19 infection patients had partial recovery of lung function, while a significant recovery of lung function was observed three months after the COVID-19 infection.
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1. Previous studies suggest that exogenous nitric oxide (NO) and NO-dependent signalling pathways modulate intracellular pH (pH(i)) in different cell types, but the role of NO in pH(i) regulation in the heart is poorly understood. Therefore, in the present study we investigated the effect of the NO donors S-nitroso-N-acetyl-DL-penicillamine, spermine NONOate and propylamine propylamine NONOate on pH(i) in rat isolated ventricular myocytes. 2. Cells were isolated from the hearts of adult Wistar rats and pH(i) was monitored using the pH-sensitive fluorescent indicator 5-(and-6)-carboxy seminaphtharhodafluor (SNARF)-1 (10 µmol/L) and a confocal microscope. To test the effect of NO donors on the Naâº/H⺠exchanger (NHE), basal pH(i) in Naâº-free buffer and pH(i) recovery from intracellular acidosis after an ammonium chloride (10 mmol/L) prepulse were monitored. The role of carbonic anhydrase was tested using acetazolamide (50 µmol/L). 4,4-Diisothiocyanatostilbene-2,2'-disulphonic acid (0.5 mmol/L; DIDS) was used to inhibit the Clâ»/OHâ» and Clâ»/HCO3-exchangers. Acetazolamide and DIDS were applied via the superfusion system 1 and 5 min before the NO donors. 3. All three NO donors acutely decreased pH(i) and this effect persisted until the NO donor was removed. In Naâº-free buffer, the decrease in basal pH(i) was increased, whereas inhibition of carbonic anhydrase and Clâ»/OHâ» and Clâ»/HCO3â» exchangers did not alter the effects of the NO donors on pH(i). After an ammonium preload, pH(i) recovery was accelerated in the presence of the NO donors. 4. In conclusion, exogenous NO decreases basal pH(i), leading to increased NHE activity. Carbonic anhydrase and chloride-dependent sarcolemmal HCO3â» and OHâ» transporters are not involved in the NO-induced decrease in pH(i) in rat isolated ventricular myocytes.
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Líquido Intracelular/metabolismo , Miocitos Cardíacos/metabolismo , Óxido Nítrico/metabolismo , Intercambiadores de Sodio-Hidrógeno/metabolismo , Cloruro de Amonio/farmacología , Animales , Antiportadores/antagonistas & inhibidores , Antiportadores/metabolismo , Inhibidores de Anhidrasa Carbónica/farmacología , Anhidrasas Carbónicas/química , Anhidrasas Carbónicas/metabolismo , Células Cultivadas , Colorantes Fluorescentes/química , Hidrazinas/farmacología , Concentración de Iones de Hidrógeno , Líquido Intracelular/efectos de los fármacos , Masculino , Moduladores del Transporte de Membrana/farmacología , Microscopía Confocal , Miocitos Cardíacos/citología , Miocitos Cardíacos/efectos de los fármacos , Óxido Nítrico/agonistas , Óxido Nítrico/farmacología , Donantes de Óxido Nítrico/farmacología , Ratas , Ratas Wistar , S-Nitroso-N-Acetilpenicilamina/farmacología , Intercambiadores de Sodio-Hidrógeno/agonistas , Intercambiadores de Sodio-Hidrógeno/antagonistas & inhibidores , Espermina/análogos & derivados , Espermina/farmacologíaRESUMEN
Painful axotomy decreases K(ATP) channel current (IK(ATP)) in primary afferent neurons. Because cytosolic Ca(2+) signaling is depressed in injured dorsal root ganglia (DRG) neurons, we investigated whether Ca(2+)-calmodulin (CaM)-Ca(2+)/CaM-dependent kinase II (CaMKII) regulates IK(ATP) in large DRG neurons. Immunohistochemistry identified the presence of K(ATP) channel subunits SUR1, SUR2, and Kir6.2 but not Kir6.1, and pCaMKII in neurofilament 200-positive DRG somata. Single-channel recordings from cell-attached patches revealed that basal and evoked IK(ATP) by ionomycin, a Ca(2+) ionophore, is activated by CaMKII. In axotomized neurons from rats made hyperalgesic by spinal nerve ligation (SNL), basal K(ATP) channel activity was decreased, and sensitivity to ionomycin was abolished. Basal and Ca(2+)-evoked K(ATP) channel activity correlated inversely with the degree of hyperalgesia induced by SNL in the rats from which the neurons were isolated. Inhibition of IK(ATP) by glybenclamide, a selective K(ATP) channel inhibitor, depolarized resting membrane potential (RMP) recorded in perforated whole-cell patches and enhanced neurotransmitter release measured by amperometry. The selective K(ATP) channel opener diazoxide hyperpolarized the RMP and attenuated neurotransmitter release. Axotomized neurons from rats made hyperalgesic by SNL lost sensitivity to the myristoylated form of autocamtide-2-related inhibitory peptide (AIPm), a pseudosubstrate blocker of CaMKII, whereas axotomized neurons from SNL animals that failed to develop hyperalgesia showed normal IK(ATP) inhibition by AIPm. AIPm also depolarized RMP in control neurons via K(ATP) channel inhibition. Unitary current conductance and sensitivity of K(ATP) channels to cytosolic ATP and ligands were preserved even after painful nerve injury, thus providing opportunities for selective therapeutic targeting against neuropathic pain.
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Proteína Quinasa Tipo 2 Dependiente de Calcio Calmodulina/metabolismo , Calcio/metabolismo , Calmodulina/metabolismo , Hiperalgesia/metabolismo , Canales KATP/metabolismo , Neuronas Aferentes/metabolismo , Animales , Axotomía , Sistema Libre de Células , Fenómenos Electrofisiológicos , Ganglios Espinales/metabolismo , Ionomicina/farmacología , Masculino , Técnicas de Placa-Clamp , Ratas , Ratas Sprague-DawleyRESUMEN
Mitochondrial bioenergetic studies mostly rely on isolated mitochondria thus excluding the regulatory role of other cellular compartments important for the overall mitochondrial function. In intact cardiomyocytes, we followed the dynamics of electron fluxes along specific sites of the electron transport chain (ETC) by simultaneous detection of NAD(P)H and flavoprotein (FP) fluorescence intensities using a laser-scanning confocal microscope. This method was used to delineate the effects of isoflurane, a volatile anesthetic and cardioprotective agent, on the ETC. Comparison to the effects of well-characterized ETC inhibitors and uncoupling agent revealed two distinct effects of isoflurane: uncoupling-induced mitochondrial depolarization and inhibition of ETC at the level of complex I. In correlation, oxygen consumption measurements in cardiomyocytes confirmed a dose-dependent, dual effect of isoflurane, and in isolated mitochondria an obstruction of the ETC primarily at the level of complex I. These effects are likely responsible for the reported mild stimulation of mitochondrial reactive oxygen species (ROS) production required for the cardioprotective effects of isoflurane. In conclusion, isoflurane exhibits complex effects on the ETC in intact cardiomyocytes, altering its electron fluxes, and thereby enhancing ROS production. The NAD(P)H-FP fluorometry is a useful method for exploring the effect of drugs on mitochondria and identifying their specific sites of action within the ETC of intact cardiomyocytes.
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Flavoproteínas/metabolismo , Isoflurano/farmacología , Mitocondrias Cardíacas/metabolismo , Miocitos Cardíacos/efectos de los fármacos , NADP/metabolismo , Anestésicos por Inhalación/farmacología , Animales , Células Cultivadas , Relación Dosis-Respuesta a Droga , Transporte de Electrón/efectos de los fármacos , Complejo I de Transporte de Electrón/metabolismo , Fluorometría/métodos , Masculino , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Microscopía Confocal , Modelos Biológicos , Miocitos Cardíacos/citología , Miocitos Cardíacos/metabolismo , Oxidación-Reducción/efectos de los fármacos , Consumo de Oxígeno/efectos de los fármacos , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismoRESUMEN
BACKGROUND: Reactive oxygen species (ROS) mediate the effects of anesthetic precondition to protect against ischemia and reperfusion injury, but the mechanisms of ROS generation remain unclear. In this study, the authors investigated if mitochondria-targeted antioxidant (mitotempol) abolishes the cardioprotective effects of anesthetic preconditioning. Further, the authors investigated the mechanism by which isoflurane alters ROS generation in isolated mitochondria and submitochondrial particles. METHODS: Rats were pretreated with 0.9% saline, 3.0 mg/kg mitotempol in the absence or presence of 30 min exposure to isoflurane. Myocardial infarction was induced by left anterior descending artery occlusion for 30 min followed by reperfusion for 2 h and infarct size measurements. Mitochondrial ROS production was determined spectrofluorometrically. The effect of isoflurane on enzymatic activity of mitochondrial respiratory complexes was also determined. RESULTS: Isoflurane reduced myocardial infarct size (40 ± 9% = mean ± SD) compared with control experiments (60 ± 4%). Mitotempol abolished the cardioprotective effects of anesthetic preconditioning (60 ± 9%). Isoflurane enhanced ROS generation in submitochondrial particles with nicotinamide adenine dinucleotide (reduced form), but not with succinate, as substrate. In intact mitochondria, isoflurane enhanced ROS production in the presence of rotenone, antimycin A, or ubiquinone when pyruvate and malate were substrates, but isoflurane attenuated ROS production when succinate was substrate. Mitochondrial respiratory experiments and electron transport chain complex assays revealed that isoflurane inhibited only complex I activity. CONCLUSIONS: The results demonstrated that isoflurane produces ROS at complex I and III of the respiratory chain via the attenuation of complex I activity. The action on complex I decreases unfavorable reverse electron flow and ROS release in myocardium during reperfusion.
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Anestésicos por Inhalación/farmacología , Transporte de Electrón/efectos de los fármacos , Precondicionamiento Isquémico Miocárdico , Isoflurano/farmacología , Mitocondrias Cardíacas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Animales , Óxidos N-Cíclicos/metabolismo , Óxidos N-Cíclicos/farmacología , Complejo I de Transporte de Electrón/metabolismo , Complejo II de Transporte de Electrones/metabolismo , Complejo III de Transporte de Electrones/metabolismo , Complejo IV de Transporte de Electrones/metabolismo , Hemodinámica/efectos de los fármacos , Técnicas In Vitro , Masculino , Mitocondrias Cardíacas/efectos de los fármacos , Infarto del Miocardio/tratamiento farmacológico , Infarto del Miocardio/patología , Reperfusión Miocárdica , Consumo de Oxígeno/efectos de los fármacos , Ratas , Ratas Wistar , Rotenona/farmacología , Marcadores de Spin , Superóxido Dismutasa/metabolismo , Desacopladores/farmacologíaRESUMEN
Distant abscesses are uncommon during the episode of acute pancreatitis (AP). However, these are possible sequalae of necrotizing pancreatitis and should be treated appropriately to prevent serious septic complications. We demonstrate a case of a 56-year-old male patient who presented with severe necrotizing pancreatitis and distant retroperitoneal abscess that reached femoral region and was detected on diagnostic imaging scans. Combination of surgical and supportive therapy was employed, and the patient recovered well with no permanent consequences. Our article highlights the importance of quick and accurate diagnosis and timely intervention in this rare type of pancreatitis complication.
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Absceso Abdominal/etiología , Absceso Abdominal/terapia , Pancreatitis Aguda Necrotizante/complicaciones , Absceso Abdominal/diagnóstico por imagen , Terapia Combinada , Drenaje , Humanos , Masculino , Persona de Mediana Edad , Espacio Retroperitoneal , Muslo/diagnóstico por imagenRESUMEN
During reperfusion, the interplay between excess reactive oxygen species (ROS) production, mitochondrial Ca(2+) overload, and mitochondrial permeability transition pore (mPTP) opening, as the crucial mechanism of cardiomyocyte injury, remains intriguing. Here, we investigated whether an induction of a partial decrease in mitochondrial membrane potential (DeltaPsi(m)) is an underlying mechanism of protection by anesthetic-induced preconditioning (APC) with isoflurane, specifically addressing the interplay between ROS, Ca(2+), and mPTP opening. The magnitude of APC-induced decrease in DeltaPsi(m) was mimicked with the protonophore 2,4-dinitrophenol (DNP), and the addition of pyruvate was used to reverse APC- and DNP-induced decrease in DeltaPsi(m). In cardiomyocytes, DeltaPsi(m), ROS, mPTP opening, and cytosolic and mitochondrial Ca(2+) were measured using confocal microscope, and cardiomyocyte survival was assessed by Trypan blue exclusion. In isolated cardiac mitochondria, antimycin A-induced ROS production and Ca(2+) uptake were determined spectrofluorometrically. In cells exposed to oxidative stress, APC and DNP increased cell survival, delayed mPTP opening, and attenuated ROS production, which was reversed by mitochondrial repolarization with pyruvate. In isolated mitochondria, depolarization by APC and DNP attenuated ROS production, but not Ca(2+) uptake. However, in stressed cardiomyocytes, a similar decrease in DeltaPsi(m) attenuated both cytosolic and mitochondrial Ca(2+) accumulation. In conclusion, a partial decrease in DeltaPsi(m) underlies cardioprotective effects of APC by attenuating excess ROS production, resulting in a delay in mPTP opening and an increase in cell survival. Such decrease in DeltaPsi(m) primarily attenuates mitochondrial ROS production, with consequential decrease in mitochondrial Ca(2+) uptake.
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Calcio/fisiología , Isoflurano/farmacología , Potencial de la Membrana Mitocondrial/fisiología , Mitocondrias Cardíacas/metabolismo , Proteínas de Transporte de Membrana Mitocondrial/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Animales , Citoprotección/efectos de los fármacos , Citoprotección/fisiología , Masculino , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias Cardíacas/efectos de los fármacos , Poro de Transición de la Permeabilidad Mitocondrial , Ratas , Ratas Wistar , Factores de TiempoRESUMEN
The sulfonylurea receptor-2 (SUR2) is a subunit of ATP-sensitive potassium channels (K(ATP)) in heart. Mice with the SUR2 gene disrupted (SUR2m) are constitutively protected from ischemia-reperfusion (I/R) cardiac injury. This was surprising because K(ATP), either sarcolemmal or mitochondrial or both, are thought to be important for cardioprotection. We hypothesized that SUR2m mice have an altered mitochondrial phenotype that protects against I/R. Mitochondrial membrane potential (ΔΨ(m)), tolerance to Ca(2+) load, and reactive oxygen species (ROS) generation were studied by fluorescence-based assays, and volumetric changes in response to K(+) were measured by light scattering in isolated mitochondria. For resting SUR2m mitochondria compared with wild type, the ΔΨ(m) was less polarized (46.1 ± 0.4 vs. 51.9 ± 0.6%), tolerance to Ca(2+) loading was increased (163 ± 2 vs. 116 ± 2 µM), and ROS generation was enhanced with complex I [8.5 ± 1.2 vs. 4.9 ± 0.2 arbitrary fluorescence units (afu)/s] or complex II (351 ± 51.3 vs. 166 ± 36.2 afu/s) substrates. SUR2m mitochondria had greater swelling in K(+) medium (30.2 ± 3.1%) compared with wild type (14.5 ± 0.6%), indicating greater K(+) influx. Additionally, ΔΨ(m) decreased and swelling increased in the absence of ATP in SUR2m, but the sensitivity to ATP was less compared with wild type. When the mitochondria were subjected to hypoxia-reoxygenation, the decrease in respiration rates and respiratory control index was less in SUR2m. ΔΨ(m) maintenance in the SUR2m intact myocytes was also more tolerant to metabolic inhibition. In conclusion, the cardioprotection observed in the SUR2m mice is associated with a protected mitochondrial phenotype resulting from enhanced K(+) conductance that partially dissipated ΔΨ(m). These results have implications for possible SUR2 participation in mitochondrial K(ATP).
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Transportadoras de Casetes de Unión a ATP/metabolismo , Metabolismo Energético , Mitocondrias Cardíacas/metabolismo , Daño por Reperfusión Miocárdica/prevención & control , Miocitos Cardíacos/metabolismo , Canales de Potasio de Rectificación Interna/metabolismo , Canales de Potasio/metabolismo , Receptores de Droga/metabolismo , Transportadoras de Casetes de Unión a ATP/genética , Adenosina Trifosfato/metabolismo , Animales , Calcio/metabolismo , Hipoxia de la Célula , Respiración de la Célula , Genotipo , Luz , Masculino , Potencial de la Membrana Mitocondrial , Ratones , Ratones Mutantes , Dilatación Mitocondrial , Daño por Reperfusión Miocárdica/metabolismo , Fenotipo , Potasio/metabolismo , Canales de Potasio de Rectificación Interna/genética , Especies Reactivas de Oxígeno/metabolismo , Receptores de Droga/genética , Dispersión de Radiación , Espectrometría de Fluorescencia , Receptores de Sulfonilureas , Factores de TiempoRESUMEN
BACKGROUND: The role of endothelial nitric oxide synthase (eNOS) in isoflurane postconditioning (IsoPC)-elicited cardioprotection is poorly understood. The authors addressed this issue using eNOS mice. METHODS: In vivo or Langendorff-perfused mouse hearts underwent 30 min of ischemia followed by 2 h of reperfusion in the presence and absence of postconditioning produced with isoflurane 5 min before and 3 min after reperfusion. Ca+-induced mitochondrial permeability transition (MPT) pore opening was assessed in isolated mitochondria. Echocardiography was used to evaluate ventricular function. RESULTS: Postconditioning with 0.5, 1.0, and 1.5 minimum alveolar concentrations of isoflurane decreased infarct size from 56 +/- 10% (n = 10) in control to 48 +/- 10%, 41 +/- 8% (n = 8, P < 0.05), and 38 +/- 10% (n = 8, P < 0.05), respectively, and improved cardiac function in wild-type mice. Improvement in cardiac function by IsoPC was blocked by N-nitro-L-arginine methyl ester (a nonselective nitric oxide synthase inhibitor) administered either before ischemia or at the onset of reperfusion. Mitochondria isolated from postconditioned hearts required significantly higher in vitro Ca+ loading than did controls (78 +/- 29 microm vs. 40 +/- 25 microm CaCl2 per milligram of protein, n = 10, P < 0.05) to open the MPT pore. Hearts from eNOS mice displayed no marked differences in infarct size, cardiac function, and sensitivity of MPT pore to Ca+, compared with wild-type hearts. However, IsoPC failed to alter infarct size, cardiac function, or the amount of Ca+ necessary to open the MPT pore in mitochondria isolated from the eNOS hearts compared with control hearts. CONCLUSIONS: IsoPC protects mouse hearts from reperfusion injury by preventing MPT pore opening in an eNOS-dependent manner. Nitric oxide functions as both a trigger and a mediator of cardioprotection produced by IsoPC.
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Anestésicos por Inhalación/farmacología , Cardiotónicos , Isoflurano/farmacología , Mitocondrias Cardíacas/efectos de los fármacos , Daño por Reperfusión Miocárdica/prevención & control , Óxido Nítrico Sintasa de Tipo III/fisiología , Permeabilidad/efectos de los fármacos , Animales , Ecocardiografía , Frecuencia Cardíaca/efectos de los fármacos , Técnicas In Vitro , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Proteínas de Transporte de Membrana Mitocondrial/efectos de los fármacos , Poro de Transición de la Permeabilidad Mitocondrial , Óxido Nítrico/fisiología , Óxido Nítrico Sintasa de Tipo III/genéticaRESUMEN
BACKGROUND: Cardioprotection by volatile anesthetic-induced preconditioning (APC) involves activation of protein kinase C (PKC). This study investigated the importance of APC-activated PKC in delaying mitochondrial permeability transition pore (mPTP) opening. METHODS: Rat ventricular myocytes were exposed to isoflurane in the presence or absence of nonselective PKC inhibitor chelerythrine or isoform-specific inhibitors of PKC-delta (rottlerin) and PKC-epsilon (myristoylated PKC-epsilon V1-2 peptide), and the mPTP opening time was measured by using confocal microscopy. Ca-induced mPTP opening was measured in mitochondria isolated from rats exposed to isoflurane in the presence and absence of chelerythrine or in mitochondria directly treated with isoflurane after isolation. Translocation of PKC-epsilon was assessed in APC and control cardiomyocytes by Western blotting. RESULTS: In cardiomyocytes, APC prolonged time necessary to induce mPTP opening (261 +/- 26 s APC vs. 216 +/- 27 s control; P < 0.05), and chelerythrine abolished this delay to 213 +/- 22 s. The effect of isoflurane was also abolished when PKC-epsilon inhibitor was applied (210 +/- 22 s) but not in the presence of PKC-delta inhibitor (269 +/- 31 s). Western blotting revealed translocation of PKC-epsilon toward mitochondria in APC cells. The Ca concentration required for mPTP opening was significantly higher in mitochondria from APC rats (45 +/- 8 microM x mg control vs. 64 +/- 8 microM x mg APC), and APC effect was reversed with chelerythrine. In contrast, isoflurane did not protect directly treated mitochondria. CONCLUSION: APC induces delay of mPTP opening through PKC-epsilon mediated inhibition of mPTP opening, but not through PKC-delta. These results point to the connection between cytosolic and mitochondrial components of cardioprotection by isoflurane.
Asunto(s)
Anestésicos/farmacología , Precondicionamiento Isquémico Miocárdico , Mitocondrias Cardíacas/fisiología , Proteínas de Transporte de Membrana Mitocondrial/efectos de los fármacos , Proteína Quinasa C-epsilon/fisiología , Transducción de Señal/fisiología , Animales , Western Blotting , Inhibidores Enzimáticos/farmacología , Procesamiento de Imagen Asistido por Computador , Técnicas In Vitro , Masculino , Microscopía Confocal , Mitocondrias Cardíacas/efectos de los fármacos , Poro de Transición de la Permeabilidad Mitocondrial , Miocitos Cardíacos/efectos de los fármacos , Permeabilidad/efectos de los fármacos , Proteína Quinasa C-delta/antagonistas & inhibidores , Proteína Quinasa C-delta/fisiología , Proteína Quinasa C-epsilon/antagonistas & inhibidores , Transporte de Proteínas/efectos de los fármacos , Ratas , Ratas Wistar , Transducción de Señal/efectos de los fármacosRESUMEN
BACKGROUND: Signal transduction cascade of anesthetic-induced preconditioning has been extensively studied, yet many aspects of it remain unsolved. Here, we investigated the roles of reactive oxygen species (ROS) and mitochondrial uncoupling in cardiomyocyte preconditioning by two modern volatile anesthetics: desflurane and sevoflurane. METHODS: Adult rat ventricular cardiomyocytes were isolated enzymatically. The preconditioning potency of desflurane and sevoflurane was assessed in cell survival experiments by evaluating myocyte protection from the oxidative stress-induced cell death. ROS production and flavoprotein fluorescence, an indicator of flavoprotein oxidation and mitochondrial uncoupling, were monitored in real time by confocal microscopy. The functional aspect of enhanced ROS generation by the anesthetics was assessed in cell survival and confocal experiments using the ROS scavenger Trolox. RESULTS: Preconditioning of cardiomyocytes with desflurane or sevoflurane significantly decreased oxidative stress-induced cell death. That effect coincided with increased ROS production and increased flavoprotein oxidation detected during acute myocyte exposure to the anesthetics. Desflurane induced significantly greater ROS production and flavoprotein oxidation than sevoflurane. ROS scavenging with Trolox abrogated preconditioning potency of anesthetics and attenuated flavoprotein oxidation. CONCLUSION: Preconditioning with desflurane or sevoflurane protects isolated rat cardiomyocytes from oxidative stress-induced cell death. Scavenging of ROS abolishes the preconditioning effect of both anesthetics and attenuates anesthetic-induced mitochondrial uncoupling, suggesting a crucial role for ROS in anesthetic-induced preconditioning and implying that ROS act upstream of mitochondrial uncoupling. Desflurane exhibits greater effect on stimulation of ROS production and mitochondrial uncoupling than sevoflurane.
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Anestésicos por Inhalación/farmacología , Precondicionamiento Isquémico Miocárdico , Isoflurano/análogos & derivados , Éteres Metílicos/farmacología , Mitocondrias/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacos , Animales , Muerte Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Cromanos/farmacología , Desflurano , Flavoproteínas/metabolismo , Depuradores de Radicales Libres/farmacología , Técnicas In Vitro , Isoflurano/farmacología , Masculino , Microscopía Confocal , Mitocondrias/efectos de los fármacos , Miocitos Cardíacos/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Wistar , SevofluranoRESUMEN
We investigated the effects of different ischemia-mimetic factors on intracellular Ca2+ concentration ([Ca2+]i). Ventricular myocytes were isolated from adult Wistar rats, and [Ca2+]i was measured using fluorescent indicator fluo-4 AM by confocal microscopy. Intracellular pH was measured using c5-(and-6)-carboxy SNARF-1 AM, a dual emission pH-sensitive ionophore. Myocytes were exposed to hypoxia, extracellular acidosis (pH(o) 6.8), Na-lactate (10 mM), or to combination of those factors for 25 min. Monitoring of [Ca2+]i using fluo-4 AM fluorescent indicator revealed that [Ca2+]i accumulation increased immediately after exposing the cells to Na-lactate and extracellular acidosis, but not during cell exposure to moderate ischemia. Increase in [Ca2+]i during Na-lactate exposure decreased to control levels at the end of exposure period at extracellular pH 7.4, but not at pH 6.8. When combined, Na-lactate and acidosis had an additive effect on [Ca2+]i increase. After removal of solutions, [Ca2+]i continued to rise only when acidosis, hypoxia, and Na-lactate were applied together. Analysis of intracellular pH revealed that treatment of cells by Na-lactate and acidosis caused intracellular acidification, while short ischemia did not significantly change intracellular pH. Our experiments suggest that increase in [Ca2+]i during short hypoxia does not occur if pH(i) does not fall, while extracellular acidosis is required for sustained rise in [Ca2+]i induced by Na-lactate. Comparing to the effect of Na-lactate, extracellular acidosis induced slower [Ca2+]i elevation, accompanied with slower decrease in intracellular pH. These multiple effects of hypoxia, extracellular acidosis, and Na-lactate are likely to cause [Ca2+]i accumulation after the hypoxic stress.