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1.
Medicine (Baltimore) ; 99(28): e20934, 2020 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-32664090

RESUMO

This study aimed to investigate the myocardial protective effect of liquid sodium phosphocreatine cardiac arrest in extracorporeal circulation surgery treating infants with atrial septal defects.Eighty-four infants with atrial septal defects who required extracorporeal circulation surgery treatment at our hospital from January 2016 to June 2018 were divided into an observation group and a control group through a digitally randomized method, with 42 cases in each group. The control group adopted the conventional modified St Thomas II high potassium cold liquid crystal cardiac arrest, while the observation group adopted the liquid sodium phosphocreatine cardiac arrest.The myocardial enzyme indexes of the 2 groups 3, 6, 12, and 24 hours postoperatively were higher than before establishing the cardiopulmonary bypass and the enzyme indexes of the control group at the same time were higher than that of the observation group; adenosine triphosphate, adenosine diphosphate, and other energy levels and the postoperative recovery rate energy levels of the observation group were higher than those in the control group, the difference was statistically significant (P < .05).Liquid sodium phosphocreatine cardiac arrest used in extracorporeal circulation surgery treating infants with atrial septal defects can reduce myocardial ischemia-reperfusion injury, maintain energy supply during ischemia, strengthen the St Thomas II effect, and aid postoperative cardiac function recovery of high potassium cold liquid crystal cardiac arrest used in infants with atrial septal defects and treated with extracorporeal circulation surgery.


Assuntos
Ponte Cardiopulmonar/métodos , Cardiotônicos/farmacologia , Parada Cardíaca Induzida/métodos , Comunicação Interatrial/cirurgia , Fosfocreatina/farmacologia , Difosfato de Adenosina/metabolismo , Trifosfato de Adenosina/metabolismo , Cardiotônicos/administração & dosagem , Estudos de Casos e Controles , Pré-Escolar , Circulação Extracorpórea/métodos , Feminino , Parada Cardíaca/induzido quimicamente , Comunicação Interatrial/diagnóstico , Comunicação Interatrial/tratamento farmacológico , Humanos , Lactente , Masculino , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Miocárdio/química , Miocárdio/enzimologia , Preservação de Órgãos/métodos , Fosfocreatina/administração & dosagem , Período Pós-Operatório , Cloreto de Potássio/administração & dosagem , Cloreto de Potássio/farmacologia , Substâncias Protetoras/administração & dosagem , Recuperação de Função Fisiológica/efeitos dos fármacos
2.
Am J Physiol Lung Cell Mol Physiol ; 318(5): L943-L952, 2020 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-32233794

RESUMO

Transient receptor potential ankyrin-1 (TRPA1) is a ligand-gated cation channel that responds to endogenous and exogenous irritants. TRPA1 is expressed on multiple cell types throughout the lungs, but previous studies have primarily focused on TRPA1 stimulation of airway sensory nerves. We sought to understand the integrated physiological airway response to TRPA1 stimulation. The TRPA1 agonists allyl isothiocyanate (AITC) and cinnamaldehyde (CINN) were tested in sedated, mechanically ventilated guinea pigs in vivo. Reproducible bronchoconstrictions were induced by electrical stimulation of the vagus nerves. Animals were then treated with intravenous AITC or CINN. AITC and CINN were also tested on isolated guinea pig and mouse tracheas and postmortem human trachealis muscle strips in an organ bath. Tissues were contracted with methacholine, histamine, or potassium chloride and then treated with AITC or CINN. Some airways were pretreated with TRPA1 antagonists, the cyclooxygenase inhibitor indomethacin, the EP2 receptor antagonist PF 04418948, or tetrodotoxin. AITC and CINN blocked vagally mediated bronchoconstriction in guinea pigs. Pretreatment with indomethacin completely abolished the airway response to TRPA1 agonists. Similarly, AITC and CINN dose-dependently relaxed precontracted guinea pig, mouse, and human airways in the organ bath. AITC- and CINN-induced airway relaxation required TRPA1, prostaglandins, and PGE2 receptor activation. TRPA1-induced airway relaxation did not require epithelium or tetrodotoxin-sensitive nerves. Finally, AITC blocked airway hyperreactivity in two animal models of allergic asthma. These data demonstrate that stimulation of TRPA1 causes bronchodilation of intact airways and suggest that the TRPA1 pathway is a potential pharmacological target for bronchodilation.


Assuntos
Dinoprostona/metabolismo , Músculo Liso/metabolismo , Canal de Cátion TRPA1/genética , Traqueia/metabolismo , Acroleína/análogos & derivados , Acroleína/farmacologia , Animais , Broncoconstrição/efeitos dos fármacos , Estimulação Elétrica , Regulação da Expressão Gênica , Cobaias , Histamina/farmacologia , Humanos , Indometacina/farmacologia , Isotiocianatos/farmacologia , Masculino , Cloreto de Metacolina/farmacologia , Camundongos , Músculo Liso/efeitos dos fármacos , Técnicas de Cultura de Órgãos , Cloreto de Potássio/farmacologia , Prostaglandina-Endoperóxido Sintases/genética , Prostaglandina-Endoperóxido Sintases/metabolismo , Respiração Artificial , Transdução de Sinais , Canal de Cátion TRPA1/agonistas , Canal de Cátion TRPA1/antagonistas & inibidores , Canal de Cátion TRPA1/metabolismo , Tetrodotoxina/farmacologia , Traqueia/efeitos dos fármacos , Nervo Vago/fisiologia
3.
J Appl Oral Sci ; 28: e20190516, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32236357

RESUMO

INTRODUCTION: This study investigated the effect of a calcium hydroxide (CH) paste (CleaniCal®) containing N-2-methyl pyrrolidone (NMP) as a vehicle on Enterococcus faecalis (E. faecalis) biofilms compared with other products containing saline (Calasept Plus™) or propylene glycol (PG) (Calcipex II®). METHODOLOGY: Standardized bovine root canal specimens were used. The antibacterial effects were measured by colony-forming unit counting. The thickness of bacterial microcolonies and exopolysaccharides was assessed using confocal laser scanning microscopy. Morphological features of the biofilms were observed using field-emission scanning electron microscopy (FE-SEM). Bovine tooth blocks covered with nail polish were immersed into the vehicles and dispelling was observed. The data were analyzed using one-way analysis of variance and Tukey tests (p<0.05). RESULTS: CleaniCal® showed the highest antibacterial activity, followed by Calcipex II® (p<0.05). Moreover, NMP showed a higher antibacterial effect compared with PG (p<0.05). The thickness of bacteria and EPS in the CleaniCal® group was significantly lower than that of other materials tested (p<0.05). FE-SEM images showed the specimens treated with Calasept Plus™ were covered with biofilms, whereas the specimens treated with other medicaments were not. Notably, the specimen treated with CleaniCal® was cleaner than the one treated with Calcipex II®. Furthermore, the nail polish on the bovine tooth block immersed in NMP was completely dispelled. CONCLUSIONS: CleaniCal® performed better than Calasept Plus™ and Calcipex II® in the removal efficacy of E. faecalis biofilms. The results suggest the effect might be due to the potent dissolving effect of NMP on organic substances.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Hidróxido de Cálcio/farmacologia , Enterococcus faecalis/efeitos dos fármacos , Pirrolidinonas/farmacologia , Irrigantes do Canal Radicular/farmacologia , Análise de Variância , Animais , Cloreto de Cálcio/química , Cloreto de Cálcio/farmacologia , Hidróxido de Cálcio/química , Bovinos , Contagem de Colônia Microbiana , Combinação de Medicamentos , Teste de Materiais , Microscopia Confocal , Microscopia Eletrônica de Varredura , Cloreto de Potássio/química , Cloreto de Potássio/farmacologia , Pirrolidinonas/química , Reprodutibilidade dos Testes , Irrigantes do Canal Radicular/química , Bicarbonato de Sódio/química , Bicarbonato de Sódio/farmacologia , Cloreto de Sódio/química , Cloreto de Sódio/farmacologia , Estatísticas não Paramétricas
4.
J Cardiothorac Surg ; 15(1): 4, 2020 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-31915024

RESUMO

BACKGROUND: Because hearts in acute myocardial infarction are often prone to ischemia-reperfusion damage during cardiac surgery, we investigated the influence of intracellular crystalloid cardioplegia solution (CCP) and extracellular blood cardioplegia solution (BCP) on cardiac function, metabolism, and infarct size in a rat heart model of myocardial infarction. METHODS: Following euthanasia, the hearts of 50 rats were quickly excised, cannulated, and inserted into a blood-perfused isolated heart apparatus. A regional myocardial infarction was created in the infarction group (18 hearts) for 120 min; the control group (32 hearts) was not subjected to infarction. In each group, either Buckberg BCP or Bretschneider CCP was administered for an aortic clamping time of 90 min. Functional parameters were recorded during reperfusion: coronary blood flow, left ventricular developed pressure (LVDP) and contractility (dp/dt max). Infarct size was determined by planimetry. The results were compared between the groups using analysis of variance or parametric tests, as appropriate. RESULTS: Cardiac function after acute myocardial infarction, 90 min of cardioplegic arrest, and 90 min of reperfusion was better preserved with Buckberg BCP than with Bretschneider CCP relative to baseline (BL) values (LVDP 54 ± 11% vs. 9 ± 2.9% [p = 0.0062]; dp/dt max. 73 ± 11% vs. 23 ± 2.7% [p = 0.0001]), whereas coronary flow was similarly impaired (BCP 55 ± 15%, CCP 63 ± 17% [p = 0.99]). The infarct in BCP-treated hearts was smaller (25% of myocardium) and limited to the area of coronary artery ligation, whereas in CCP hearts the infarct was larger (48% of myocardium; p = 0.029) and myocardial necrosis was distributed unevenly to the left ventricular wall. CONCLUSIONS: In a rat model of acute myocardial infarction followed by cardioplegic arrest, application of BCP leads to better myocardial recovery than CCP.


Assuntos
Soluções Cardioplégicas/farmacologia , Soluções Cristaloides/farmacologia , Infarto do Miocárdio/cirurgia , Miocárdio/patologia , Compostos de Potássio/farmacologia , Animais , Circulação Coronária/efeitos dos fármacos , Glucose/farmacologia , Parada Cardíaca Induzida/métodos , Masculino , Manitol/farmacologia , Miocárdio/metabolismo , Necrose , Cloreto de Potássio/farmacologia , Procaína/farmacologia , Ratos , Função Ventricular Esquerda/efeitos dos fármacos
5.
PLoS One ; 15(1): e0228335, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31978138

RESUMO

Precise choice of potassium (K) source and application method does matter for its cost-effectiveness. This study was aimed to evaluate the best source and method of K fertilizer application to improve cotton productivity and profitability under an arid climate. Three different K sources (KNO3, K2SO4 and KCl) were applied at 100 kg ha-1 by four methods, i.e. a) basal application, b) side dressing, c) fertigation and d) foliar application of 2% K2SO4. The highest productivity and profitability were recorded with K2SO4 applied as foliar application. Total boll weight per plant was similar in foliar applied K2SO4 and basal application of KNO3. Better boll opening in foliar applied K2SO4, perhaps, played decisive role for increased seed-cotton yield. For basal application and side dressing, KNO3 produced the highest seed-cotton yield, but the benefit cost ratio was better for foliar applied K2SO4. In crux, foliar application of K2SO4 might be opted to improve the seed cotton yield, fiber quality and net returns under the arid climate. However, soil K application through K2SO4 and/or KNO3 is essential to balance the K removal from soil.


Assuntos
Misturas Complexas/química , Gossypium/crescimento & desenvolvimento , Nitratos/farmacologia , Cloreto de Potássio/farmacologia , Compostos de Potássio/farmacologia , Sulfatos/farmacologia , Clima Desértico , Fertilizantes , Gossypium/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Sementes/efeitos dos fármacos , Sementes/crescimento & desenvolvimento , Solo/química
6.
Ann Thorac Surg ; 109(3): 763-770, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31470011

RESUMO

BACKGROUND: Various solutions are used for donor heart preservation. We examined the outcomes in our heart transplant population where histidine-tryptophan-ketoglutarate (HTK) solution has been used for heart preservation since 2004. METHODS: This was a retrospective review of the United Network for Organ Sharing (UNOS) database (2004-2016) comparing our heart transplant outcomes with other national centers. Propensity matching in a 1:3 ratio was performed to adjust for preoperative recipient variables. RESULTS: After propensity matching comparing UNOS outcomes (n = 1080) with our institutional data (n = 360), there was no difference in matched preoperative variables. Donor hearts were similar for donor age, sex, donor-to-recipient size ratio, LVEF, and ischemic time. Our HTK cohort had a larger proportion with donor cardiac arrest (26.3% vs 6.1%, P < .001) and longer cardiac arrest duration (22.1 ± 16.0 vs 17.2 ± 14.0 minutes, P = .052). Our primary graft dysfunction (PGD) rate requiring mechanical support was 4.2% (n = 1). Postoperative mechanical support use for PGD included extracorporeal membrane oxygenation in 9 (60.0%), intraaortic balloon pump in 4 (26.7%), right ventricular assist device in 3 (20%), and biventricular assist device in 3 (20%). Overall survival at our institution was similar to the national average (P = .649). Survival at 1, 5, and 10 years with HTK was 92.2%, 81.3%, and 70.8%, and for the UNOS population was 91.6%, 80.3%, and 62.0%, respectively. CONCLUSIONS: Use of HTK solution for donor hearts was associated with a low rate of severe PGD. Overall survival was not significantly different from other institutions using a variety of preservation solutions in the UNOS database during the same period. HTK solution is efficacious for preservation of donor hearts.


Assuntos
Transplante de Coração/métodos , Preservação de Órgãos/métodos , Disfunção Primária do Enxerto/prevenção & controle , Doadores de Tecidos , Feminino , Glucose/farmacologia , Sobrevivência de Enxerto , Humanos , Incidência , Masculino , Manitol/farmacologia , Michigan/epidemiologia , Pessoa de Meia-Idade , Soluções para Preservação de Órgãos/farmacologia , Cloreto de Potássio/farmacologia , Disfunção Primária do Enxerto/epidemiologia , Procaína/farmacologia , Estudos Retrospectivos , Taxa de Sobrevida/tendências , Fatores de Tempo
7.
Interact Cardiovasc Thorac Surg ; 30(1): 136-143, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-31873745

RESUMO

OBJECTIVES: The optimal myocardial protective solution in the neonatal arterial switch operation remains controversial. The aim of this study was to demonstrate that Bretschneider's histidine-tryptophan-ketoglutarate crystalloid solution (Custodiol) offers protection at least similar to that of cold blood cardioplegia. METHODS: Patients who underwent the neonatal arterial switch operation with Custodiol between January 2016 and December 2018 (n = 23) were compared with an historical cohort from August 2010 to December 2015 in which cold blood cardioplegia was used (n = 41). A linear mixed-effect model for repeated measures was performed to test the recovery of myocardial function based on inotropic and vasoactive inotropic scores, cardiac enzyme release and left ventricular ejection fraction. RESULTS: Patients in the cold blood cardioplegia group had higher inotropic scores in the first 24 h (0 h, P = 0.001 and 24 h, P = 0.006) and higher vasoactive inotropic scores in the first 72 h (0 h, 24 h and 48 h, P < 0.001; 72 h, P = 0.012). Cardiac troponin-I concentrations were higher in the cold blood cardioplegia group at postoperative hours 1-72 (1 h, 6 h, 12 h and 24 h, P < 0.001; 48 h, P = 0.001 and 72 h, P = 0.003). Creatinine-kinase-MB concentrations were higher in the cold blood cardioplegia group at postoperative hours 1-24 (1 h, 6 h and 12 h, P < 0.001; 24 h, P = 0.042). The left ventricular ejection fraction was higher in the Custodiol group just after the operation (P = 0.005), at 24 h (P = 0.001) and on the first day without inotropic support (P = 0.011). CONCLUSIONS: Neonatal myocardium protected with Custodiol during the arterial switch operation presented optimal ventricular function recovery with less inotropic support and less myocardial damage compared with cold blood cardioplegia.


Assuntos
Transposição das Grandes Artérias/métodos , Parada Cardíaca Induzida/métodos , Miocárdio/metabolismo , Transposição dos Grandes Vasos/cirurgia , Soluções Cardioplégicas/farmacologia , Feminino , Glucose/farmacologia , Humanos , Recém-Nascido , Masculino , Manitol/farmacologia , Cloreto de Potássio/farmacologia , Procaína/farmacologia , Transposição dos Grandes Vasos/sangue , Troponina I/sangue , Função Ventricular Esquerda/efeitos dos fármacos
8.
J Thorac Cardiovasc Surg ; 158(6): 1543-1554.e8, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31753163

RESUMO

OBJECTIVE: Cardiac surgery demands highly effective cardioprotective regimens. We previously demonstrated improved cardioprotection with "polarized" compared with "depolarized" arrest. This study uses a clinically relevant porcine model of cardiopulmonary bypass to compare the efficacy of blood-based St Thomas' Hospital polarizing cardioplegia (STH-Pol-B) with blood-based St Thomas' Hospital hyperkalemic cardioplegia (STH2-B). METHODS: Pigs were monitored and subjected to normothermic cardiopulmonary bypass, cardiac arrest via antegrade cold (4°C) blood cardioplegia (STH2-B, control group: n = 6 or STH-Pol-B, study group: n = 7), and global ischemia (60 minutes) followed by on-pump reperfusion (60 minutes) and subsequent off-pump reperfusion (90 minutes). At termination, tissue samples were taken for analysis of high-energy phosphates, ultrastructure, and microRNAs. The primary endpoint of this study was creatine kinase-muscle/brain release during reperfusion. RESULTS: Creatine kinase-muscle/brain was comparable in both groups. After pigs were weaned from cardiopulmonary bypass, hemodynamic parameters such as mean arterial pressure (P = .007), left ventricular systolic pressure (P < .001), external heart work (P = .012), stroke volume (P = .015), as well as dp/dtmax (P = .027), were improved with polarizing cardioplegia. Wedge pressure was significantly lower in the study group (P < .01). Energy charge was comparable between groups. MicroRNA-708-5p was significantly lower (P = .019) and microRNA-122 expression significantly (P = .046) greater in STH-Pol-B hearts. CONCLUSIONS: Polarized cardiac arrest offers similar myocardial protection and enhances functional recovery in a porcine model of cardiopulmonary bypass. Differential expression of microRNAs may indicate possible new ischemia-reperfusion markers. These results confirm the noninferiority and potential of polarized versus depolarized arrest.


Assuntos
Soluções Cardioplégicas/farmacologia , Ponte Cardiopulmonar , Parada Circulatória Induzida por Hipotermia Profunda , Hemodinâmica/efeitos dos fármacos , Função Ventricular Esquerda/efeitos dos fármacos , Animais , Bicarbonatos/farmacologia , Biomarcadores/sangue , Cloreto de Cálcio/farmacologia , Ponte Cardiopulmonar/efeitos adversos , Parada Circulatória Induzida por Hipotermia Profunda/efeitos adversos , Creatina Quinase Forma MB/sangue , Metabolismo Energético/efeitos dos fármacos , Feminino , Magnésio/farmacologia , MicroRNAs/metabolismo , Modelos Animais , Miocárdio/metabolismo , Miocárdio/patologia , Cloreto de Potássio/farmacologia , Recuperação de Função Fisiológica , Cloreto de Sódio/farmacologia , Sus scrofa , Fatores de Tempo
9.
Exp Mol Pathol ; 111: 104318, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31614130

RESUMO

Ketamine is widely used both as anesthetic and abuse drug. In this study, we investigated the effects of a wide range of ketamine concentrations (100-500-1000 µM) on calcium mobilization and the induction of cell death in undifferentiated PC12 cells, 24 h after treatment. Calcium mobilization was measured as the percentage of fluorescence one minute after depolarization by flow cytometry. For the kinetic changes in [Ca2+]c, fluorescence microscopy with Live Imaging was used with a resolution time of 0.87 s (exposure time: 20 ms). Fluo-4 AM was used for both methods. Flow cytometry using TMRE, NAO, and Annexin V-FITC/PI probes were employed for the evaluation of mitochondrial membrane potential (ΔΨm), cardiolipin content and type of cell death respectively. Fluorescence microscopy was used for the evaluation of DNA fragmentation by TUNEL assay with dUTP-conjugated FITC. Results obtained by flow cytometry showed a clear increment in cell response to depolarization after addition of 50 mM and 70 mM KCl in PC12 cells. Simultaneously, cells treated with 100 µM and 500 µM ketamine during 24 h, induced a decreased response to depolarization as compared with control cells. In addition, 1000 µM ketamine induced a similar increase in Fluo4AM fluorescence either after addition of 50 or 70 mM KCl. The kinetic assays showed that after 100 mM KCl, cells pre-treated with ketamine showed a marked decrease in [Ca2+]c as compared with control cells. In the case of 1000 µM ketamine treatment, an increased and sustained [Ca2+]c was observed along the whole assay, indicating a cell disability to maintain calcium homeostasis. Associated with these cytosolic calcium alterations, mitochondrial depolarization, cardiolipin depletion and alteration in Bax protein expression were observed after ketamine treatment. Our data demonstrate that ketamine action in these cells seems to be independent from NMDAR, as observed by the absence of glutamate­calcium response. Acute disturbance in [Ca2+]c could be mediated by the inhibition of VDCCs as part of the molecular mechanism of ketamine cytotoxicity leading to mitochondrial dysfunction and cell death by apoptosis and necrosis.


Assuntos
Canais de Cálcio/metabolismo , Ketamina/farmacologia , Analgésicos/administração & dosagem , Analgésicos/farmacologia , Animais , Sinalização do Cálcio/efeitos dos fármacos , Cardiolipinas/metabolismo , Morte Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ketamina/administração & dosagem , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Células PC12 , Cloreto de Potássio/farmacologia , Ratos , Proteína X Associada a bcl-2/metabolismo
10.
PLoS One ; 14(10): e0222683, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31574082

RESUMO

Mesenchymal stem cells (MSCs) were obtained from human bone marrow and amplified in cultures supplemented with human platelet lysate in order to generate myofibroblasts. When MSCs were seeded in solid collagen scaffolds, they differentiated into myofibroblasts that were observed to strongly bind to the substrate, forming a 3D cell scaffold network that developed tension and shortening after KCl stimulation. Moreover, MSC-laden scaffolds recapitulated the Frank-Starling mechanism so that active tension increased in response to increases in the initial length of the contractile system. This constituted a bioengineering tissue that exhibited the contractile properties observed in both striated and smooth muscles. By using the A. F. Huxley formalism, we determined the myosin crossbridge (CB) kinetics of attachment (f1) and detachment (g1 and g2), maximum myosin ATPase activity, molar myosin concentration, unitary CB force and maximum CB efficiency. CB kinetics were dramatically slow, characterizing the non-muscle myosin type IIA (NMMIIA) present in myofibroblasts. When MSCs were seeded in solid collagen scaffolds functionalized with Arg-Gly-Asp (RGD), contractility increased and CB kinetics were modified, whereas the unitary NMMIIA-CB force and maximum CB efficiency did not change. In conclusion, we provided a non-muscle bioengineering tissue whose molecular mechanical characteristics of NMMIIA were very close to those of a non-muscle contractile tissue such as the human placenta.


Assuntos
Músculo Liso/metabolismo , Cadeias Pesadas de Miosina/química , Oligopeptídeos/metabolismo , Peptídeos/metabolismo , Plaquetas/metabolismo , Células da Medula Óssea/metabolismo , Diferenciação Celular/genética , Colágeno/química , Colágeno/metabolismo , Humanos , Cinética , Células-Tronco Mesenquimais/metabolismo , Contração Muscular/genética , Miofibroblastos/metabolismo , Cadeias Pesadas de Miosina/genética , Miosinas/química , Miosinas/metabolismo , Oligopeptídeos/química , Peptídeos/química , Cloreto de Potássio/farmacologia
11.
Iran J Allergy Asthma Immunol ; 18(3): 320-331, 2019 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-31522439

RESUMO

In this study we aimed to examine the relaxant effect of berberine, a compound extracted from a variety of herbs, on rat tracheal smooth muscle (TSM) and its possible mechanism(s). Cumulative concentrations of berberine (20, 65, 200 and 600 µg/mL) were added on pre-contracted TSM by methacholine or KCl in non-incubated or incubated tissues with atropine, chlorpheniramine, propranolol, diltiazem, glibenclamide, indomethacin, L-NG-nitro arginine methyl ester (L-NAME) and papaverine. The relaxant effects of theophylline (0.2, 0.4, 0.6 and 0.8 mM) as positive control and saline (1 mL) as negative control were also examined in non-incubated tissues. Berberine showed significant and concentration-dependent relaxant effects in non-incubated tissues contracted by KCl and methacholine (p<0.01 to p<0.001). There was no significant difference in the relaxant effects of berberine between non-incubated and incubated tissues with atropine, propranolol, diltiazem, glibenclamide, and papaverine. The relaxant effects of second concentrations of berberine in incubated tissues with L-NAME, its three lower concentration in incubated tissues with chlorpheniramine and its all concentrations in incubated tissues with indomethacin were significantly lower than non-incubated tissues (p<0.05 to p<0.001). The EC50 values of berberine in incubated tissues with chlorpheniramine was significantly higher than the non-incubated condition (p<0.05). Our findings reveal a relatively potent relaxant effect of berberine that is lower than the effect of theophylline. Proposed mechanisms for the relaxant effect of berberine are histamine (H1) receptor blockade, inhibition of cyclooxygenase pathways and/or nitric oxide formation.


Assuntos
Berberina/farmacologia , Músculo Liso/efeitos dos fármacos , Músculo Liso/metabolismo , Óxido Nítrico/metabolismo , Prostaglandina-Endoperóxido Sintases/metabolismo , Receptores Histamínicos H1/metabolismo , Transdução de Sinais , Animais , Berberina/química , Broncodilatadores/química , Broncodilatadores/farmacologia , Relação Dose-Resposta a Droga , Feminino , Masculino , Modelos Biológicos , Cloreto de Potássio/farmacologia , Ratos , Traqueia/efeitos dos fármacos , Traqueia/metabolismo
12.
Am J Physiol Renal Physiol ; 317(5): F1132-F1141, 2019 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-31432708

RESUMO

Voltage-dependent L-type Ca2+ channels (L-VDCCs) and the RhoA/Rho kinase pathway are two predominant intracellular signaling pathways that regulate renal microvascular reactivity. Traditionally, these two pathways have been thought to act independently; however, recent evidence suggests that these pathways could be convergent. We hypothesized that Rho kinase inhibitors can influence L-VDCC signaling. The effects of Rho kinase inhibitors Y-27632 or RKI-1447 on KCl-induced depolarization or the L-VDCC agonist Bay K8644 were assessed in afferent arterioles using an in vitro blood-perfused rat juxtamedullary nephron preparation. Superfusion of KCl (30-90 mM) led to concentration-dependent vasoconstriction of afferent arterioles. Administration of Y-27632 (1, 5, and 10 µM) or RKI-1447 (0.1, 1, and 10 µM) significantly increased the starting diameter by 16-65%. KCl-induced vasoconstriction was markedly attenuated with 5 and 10 µM Y-27632 and with 10 µM RKI-1447 (P < 0.05 vs. KCl alone). Y-27632 (5 µM) also significantly attenuated Bay K8644-induced vasoconstriction (P < 0.05). Changes in intracellular Ca2+ concentration ([Ca2+]i) were estimated by fura-2 fluorescence during KCl-induced depolarization in cultured A7r5 cells and in freshly isolated preglomerular microvascular smooth muscle cells. Administration of 90 mM KCl significantly increased fura-2 fluorescence in both cell types. KCl-mediated elevation of [Ca2+]i in A7r5 cells was suppressed by 1-10 µM Y-27632 (P < 0.05), but 10 µM Y-27632 was required to suppress Ca2+ responses in preglomerular microvascular smooth muscle cells. RKI-1447, however, significantly attenuated KCl-mediated elevation of [Ca2+]i. Y-27632 markedly inhibited Bay K8644-induced elevation of [Ca2+]i in both cell types. The results of the present study indicate that the Rho kinase inhibitors Y-27632 and RKI-1447 can partially inhibit L-VDCC function and participate in L-VDCC signaling.


Assuntos
Aorta/citologia , Canais de Cálcio/metabolismo , Rim/irrigação sanguínea , Miócitos de Músculo Liso/efeitos dos fármacos , Transdução de Sinais/fisiologia , Quinases Associadas a rho/antagonistas & inibidores , Éster Metílico do Ácido 3-Piridinacarboxílico, 1,4-Di-Hidro-2,6-Dimetil-5-Nitro-4-(2-(Trifluormetil)fenil)/farmacologia , Amidas/farmacologia , Animais , Arteríolas/efeitos dos fármacos , Proteínas de Bactérias , Linhagem Celular , Masculino , Miócitos de Músculo Liso/metabolismo , Cloreto de Potássio/farmacologia , Piridinas/farmacologia , Ratos , Proteínas Repressoras , Tiazóis/farmacologia , Ureia/análogos & derivados , Ureia/farmacologia , Vasoconstrição/efeitos dos fármacos
13.
Food Microbiol ; 84: 103240, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31421790

RESUMO

This study evaluated whether the pre-exposure (24, 48 and 72 h) to sublethal conditions caused by acetic acid (AA), lactic acid (LA), sodium chloride (NaCl) or potassium chloride (KCl) could induce increased cross-tolerance to the essential oils from Origanum vulgare L. (OVEO) and Rosmarinus officinalis L. (ROEO) in different Listeria monocytogenes strains. Damage to membrane integrity, membrane potential, enzymatic activity and efflux activity in L. monocytogenes cells pre-exposed (24 h) to AA or NaCl and further treated with OVEO or ROEO (8 and 24 h) were investigated using flow cytometry (FC). Results of minimum inhibitory concentration (MIC) modulation test showed that pre-exposure to sublethal conditions caused by organic acids or salts increased cross-tolerance only to ROEO, since MIC of ROEO increased up to 4.8-fold against pre-exposed cells. Otherwise, MIC of OVEO against these pre-exposed cells was up to ten-fold lower than that observed against not pre-exposed cells, indicating no increase in cross-tolerance. Bacterial survival assays showed that ROEO only decreased the counts over time of cells not pre-exposed to organic acids or salts, while OVEO decreased similarly or more the counts of pre-exposed cells compared to not pre-exposed cells. Results of FC analysis showed that all measured functions in L. monocytogenes cells pre-exposed to AA or NaCl and treated with OVEO or ROEO were affected, although with different intensities. These data indicate that exposure to sublethal conditions imposed by organic acids or salts could result in a phenotype of increased cross-tolerance to ROEO but not to OVEO in L. monocytogenes.


Assuntos
Listeria monocytogenes/efeitos dos fármacos , Óleos Voláteis/farmacologia , Origanum/química , Rosmarinus/química , Sais/farmacologia , Ácido Acético/farmacologia , Cicloexanóis/farmacologia , Microbiologia de Alimentos , Conservantes de Alimentos/farmacologia , Listeria monocytogenes/fisiologia , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Cloreto de Potássio/farmacologia , Cloreto de Sódio/farmacologia , Estresse Fisiológico
14.
Mol Syst Biol ; 15(8): e8939, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31464369

RESUMO

Cells respond to environmental fluctuations by regulating multiple transcriptional programs. This response can be studied by measuring the effect of environmental changes on the transcriptome or the proteome of the cell at the end of the response. However, the dynamics of the response reflect the working of the regulatory mechanisms in action. Here, we utilized a fluorescent stress reporter gene to track the dynamics of protein production in yeast responding to environmental stress. The response is modulated by changes in both the duration and rate of transcription. We probed the underlying molecular pathways controlling these two dimensions using a library of ~1,600 single- and double-mutant strains. Dissection of the effects of these mutants and the interactions between them identified distinct modulators of response duration and response rate. Using a combination of mRNA-seq and live-cell microscopy, we uncover mechanisms by which Msn2/4, Mck1, Msn5, and the cAMP/PKA pathway modulate the response of a large module of stress-induced genes in two discrete regulatory phases. Our results and analysis show that transcriptional stress response is regulated by multiple mechanisms that overlap in time and cellular location.


Assuntos
Proteínas de Ligação a DNA/genética , Regulação Fúngica da Expressão Gênica , Quinase 3 da Glicogênio Sintase/genética , RNA Mensageiro/genética , Proteínas de Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/genética , Fatores de Transcrição/genética , AMP Cíclico/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/genética , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Proteínas de Ligação a DNA/metabolismo , Perfilação da Expressão Gênica , Interação Gene-Ambiente , Genes Reporter , Quinase 3 da Glicogênio Sintase/deficiência , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Carioferinas/genética , Carioferinas/metabolismo , Mutação , Cloreto de Potássio/farmacologia , RNA Mensageiro/metabolismo , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Transdução de Sinais , Estresse Fisiológico , Fatores de Transcrição/metabolismo , Transcrição Genética
15.
J Card Surg ; 34(10): 969-975, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31332833

RESUMO

OBJECTIVES: Cold crystalloid cardioplegia for donor heart harvesting and cold ischemic storage conditions during the transportation is the standard of care during heart transplantation procedure. Organ care system (OCS) was introduced for more prolonged and reliable ex vivo organ management. This study evaluated the two different techniques used for myocardial preservation during the procurement and transportation of the heart using the OCS. METHODS: We performed prospective analysis of 43 patients with heart failure undergoing heart transplantation and using the OCS for donor organ transport. Donor hearts were arrested using blood cardioplegia and conditioning (n = 30) or standard Custodiol (SC) solution ( n = 13). Perfusion and cardiac function parameters were continuously monitored while the donor hearts were perfused in the OCS. Impact of preservation techniques on biochemical parameters and clinical outcomes were evaluated. RESULTS: All donor hearts had stable perfusion and lactate characteristics in the OCS, with similar measures between the two groups at the beginning of the ex vivo perfusion. Ex vivo heart perfusion mean ending concentration of Interleukin (IL)-6 and IL-8 was significantly lower in the blood cardioplegia group compared to the standard care group. Clinical outcomes were comparable between the two groups of patients. CONCLUSIONS: The use of blood cardioplegia and conditioning could be a safe method for myocardial protection in distant procurement and preservation of donor hearts in the OCS.


Assuntos
Parada Cardíaca Induzida/métodos , Insuficiência Cardíaca/cirurgia , Transplante de Coração/métodos , Preservação de Órgãos/métodos , Perfusão/métodos , Doadores de Tecidos , Adulto , Feminino , Seguimentos , Glucose/farmacologia , Humanos , Masculino , Manitol/farmacologia , Soluções para Preservação de Órgãos/farmacologia , Cloreto de Potássio/farmacologia , Procaína/farmacologia , Estudos Prospectivos
16.
J Zoo Wildl Med ; 50(1): 123-126, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-31120670

RESUMO

Immersion euthanasia methods reported over the most recent decades for aquatic invertebrates use organic alcohols or halogenated hydrocarbons that can interfere with nuclear magnetic resonance (NMR) analysis. A rolling study design evaluated potassium chloride (KCl), magnesium chloride (MgCl2), and magnesium sulfate (MgSO4) as potential ion-based euthanasia methods for moon jellyfish (Aurelia aurita) destined for metabolomic analysis by NMR spectroscopy. Death was defined as the cessation of autonomous bell pulsing and response to external stimulus. MgCl2 applied at a dose of 142 g/L provided euthanasia within 32 sec of applications without the untoward effects observed with the other two salts. Euthanasia with KCl at the doses tested was associated with abnormal behavior and tissue degradation during dissection. MgSO4 at the doses tested resulted in abnormal behavior and failed to provide rapid euthanasia.


Assuntos
Eutanásia Animal/métodos , Cloreto de Magnésio/administração & dosagem , Sulfato de Magnésio/administração & dosagem , Cloreto de Potássio/administração & dosagem , Cifozoários/efeitos dos fármacos , Animais , Íons/administração & dosagem , Íons/farmacologia , Cloreto de Magnésio/farmacologia , Sulfato de Magnésio/farmacologia , Cloreto de Potássio/farmacologia , Cifozoários/fisiologia
17.
Acta Cir Bras ; 34(4): e201900402, 2019 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-31038582

RESUMO

PURPOSE: To evaluate the effect of amniotic fluid in liver preservation in organ transplantation, and compare it with standard preservation solutions. METHODS: The groups consisted of Group 1: Ringer Lactate (RL) group, Group 2: HTK group, Group 3: UW group, Group 4: AF group. The livers of rats from Group 1, 2, 3, and 4 were perfused and placed into falcon tubes containing RL, HTK, UW, and AF solutions at +4 °C, respectively. The tubes were stored for 12 hours in the refrigerator at +4°C. Tissue samples were taken at the 6th and 12th hours for histopathological examinations of the perfused livers, and storage solutions for biochemical analyzes at 6th and 12th hours. RESULTS: AF was shown to maintain organ viability by reducing the number of cells undergoing apoptosis. Histopathological changes such as sinusoidal dilatation, hydropic degeneration, and focal necrosis were found to be similar to the groups in which the standard organ preservation solutions were used. Additionally, the results of INOS, IL-10, and TNF-α,which were evaluated immunohistochemically, have been shown to be similar to the UW and HTK groups. CONCLUSIONS: AF provided conservation similar to UW and HTK in the 12-hour liver SCS process. The fact that apoptosis values are comparable to standard preservation solutions supports the success of AF in the cold storage of the liver.


Assuntos
Líquido Amniótico , Criopreservação/métodos , Fígado , Soluções para Preservação de Órgãos/farmacologia , Animais , Glucose/farmacologia , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Interleucina-10/análise , Fígado/irrigação sanguínea , Fígado/patologia , Masculino , Manitol/farmacologia , Óxido Nítrico Sintase Tipo II/análise , Preservação de Órgãos/métodos , Cloreto de Potássio/farmacologia , Procaína/farmacologia , Distribuição Aleatória , Ratos Wistar , Valores de Referência , Traumatismo por Reperfusão/prevenção & controle , Reprodutibilidade dos Testes , Solução de Ringer/farmacologia , Fatores de Tempo , Sobrevivência de Tecidos , Fator de Necrose Tumoral alfa/análise
18.
Xenotransplantation ; 26(4): e12512, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30968460

RESUMO

BACKGROUND AND AIMS: Cell-based therapies for liver disease such as bioartificial liver rely on a large quantity and high quality of hepatocytes. Cold storage was previously shown to be a better way to preserve the viability and functionality of hepatocytes during transportation rather than freezing, but this was only proved at a lower density of rat hepatocytes spheroids. The purpose of this study was to optimize conditions for cold storage of high density of primary porcine hepatocyte spheroids. METHODS: Porcine hepatocytes were isolated by a three-step perfusion method; hepatocyte spheroids were formed by a 24 hours rocked culture technique. Hepatocyte cell density was 5 × 106 /mL in 1000 mL spheroid forming medium. Spheroids were then maintained in rocked culture at 37°C (control condition) or cold stored at 4°C for 24, 48 or 72 hours in four different cold storage solutions: histidine-tryptophan-ketoglutarate (HTK) alone; HTK + 1 mM deferoxamine (DEF); HTK + 5 mM N-acetyl-L-cysteine (NAC); and HTK + 1 mM DEF + 5 mM NAC. The viability, ammonia clearance, albumin production, gene expression, and functional activity of cytochrome P450 enzymes were measured after recovery from the cold storage. RESULTS: In this study, we observed that cold-induced injury was reduced by the addition of the iron chelator. Viability of HTK + DEF group hepatocyte spheroids was increased compared with other cold storage groups (P < 0.05). Performance metrics of porcine hepatocyte spheroids cold stored for 24 hours were similar to those in control conditions. The hepatocyte spheroids in control conditions started to lose their ability to clear ammonia while production of albumin was still active at 48 and 72 hours (P < 0.05). In contrast, the viability and functionality of hepatocyte spheroids including ammonia clearance and albumin secretion were preserved in HTK + DEF group at both 48- and 72-hour time points (P < 0.05). CONCLUSIONS: The beneficial effects of HTK supplemented with DEF were more obvious after cold storage of high density of porcine hepatocyte spheroids for 72 hours. The porcine hepatocyte spheroids were above the cutoff criteria for use in a spheroid-based bioartificial liver.


Assuntos
Criopreservação/métodos , Hepatócitos/citologia , Fígado Artificial , Esferoides Celulares/citologia , Acetilcisteína/farmacologia , Albuminas/metabolismo , Amônia/metabolismo , Animais , Desferroxamina/farmacologia , Glucose/farmacologia , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Quelantes de Ferro/farmacologia , Manitol/farmacologia , Taxa de Depuração Metabólica , Soluções para Preservação de Órgãos/farmacologia , Oxirredução , Cloreto de Potássio/farmacologia , Procaína/farmacologia , Esferoides Celulares/efeitos dos fármacos , Esferoides Celulares/metabolismo , Suínos , Transplante Heterólogo
19.
Neurourol Urodyn ; 38(5): 1222-1228, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30947371

RESUMO

INTRODUCTION: Dynamic elasticity is an acutely regulated bladder material property through which filling and passive emptying produce strain softening, and active voiding restores baseline pressure. The aim of this study was to test the hypothesis that strain softening produced by filling-passive emptying is equivalent to that produced by compression-release in a porcine bladder model. METHODS/MATERIALS: Latex balloons and ex vivo perfused pig bladders were used for a series of alternating fill-passive emptying ("Fill") and external compress-release ("Press") protocols. For the Fill protocol balloons/bladders were (1) filled to defined volumes (prestrain softening), (2) filled to capacity to strain soften (reference), and (3) passively emptied to the original volume (poststrain softening). For the Press protocol, balloons/bladders were (1) filled to defined volumes (prestrain softening), (2) externally compressed to reference pressure and then released for five cycles (poststrain softening). After each protocol, bladders were voided with high-KCl buffer to induce "active" voiding. RESULTS: In both balloons and porcine bladder, both the Fill and Press protocols produced significant strain softening (P < 0.05) and poststrain softening pressures were not different for Fill and Press protocols (P > 0.05), indicating a similar degree of strain softening with both methods. CONCLUSIONS: Repeated external compression can induce bladder strain softening similar to filling and passive emptying. This technique may represent a means to acutely regulate bladder compliance and potentially be used as a mechanical treatment for urinary urgency.


Assuntos
Bexiga Urinária Hiperativa/terapia , Bexiga Urinária/patologia , Animais , Fenômenos Biomecânicos , Elasticidade , Feminino , Masculino , Cloreto de Potássio/farmacologia , Pressão , Suínos , Urodinâmica
20.
PLoS One ; 14(3): e0214336, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30893362

RESUMO

Evidence suggests that ethanol-induced hypertension is associated with increased cardiovascular responsiveness to vasopressors in vivo and enhanced reactivity of isolated arteries to vasopressors ex vivo. The underlying mechanisms are not well understood and the contribution of ethanol metabolites to vascular effects induced by ethanol consumption are unclear. Mesenteric resistance arteries were harvested from Sprague-Dawley rats. Pressure myography was utilized to test effects of ethanol, acetaldehyde and phosphatidylethanol on myogenic tone and on vasoconstriction induced by phenylephrine, arginine vasopressin (aVP), endothelin-1 and KCl. Ethanol, acetaldehyde and phosphatidylethanol concentrations were monitored during the experiments. Ethanol concentrations in the vessel bath decreased with a half-life of 25min; acetaldehyde and phosphatidylethanol concentrations remained constant. Pretreatment with ethanol dose-dependently increased the potency of phenylephrine to induce vasoconstriction 4-fold (p<0.01). These effects were comparable when arteries were pre-treated with a single dose of ethanol for 30min and when ethanol concentrations were kept constant during 30min and 60min of pretreatment. While ethanol also dose-dependently increased the potency of aVP to induce vasoconstriction 1.7-fold (p<0.05), it did not affect vasoconstriction induced by endothelin-1 or KCl. Acetaldehyde pre-treatment (30 min) dose-dependently increased the potency of phenylephrine to induce vasoconstriction 2.7-fold (p<0.01) but did not affect other vasoconstrictor responses. Phosphatidylethanol did not affect any vasoconstrictor responses. Ethanol and its metabolites did not affect myogenic tone. These data suggest that ethanol and acetaldehyde selectively sensitize intrinsic constrictor responses upon activation of vascular α1-adrenergic and/or vasopressin receptors at clinically relevant concentrations. Our findings support the concept that enhanced vasoreactivity to vasoactive hormones contributes to the development of hypertension induced by ethanol consumption. Ex vivo exposure of resistance arteries to ethanol and acetaldehyde resembles effects of chronic ethanol consumption on intrinsic vascular function, and thus could serve as test platform to evaluate interventions aimed to mitigate vascular effects associated with ethanol consumption.


Assuntos
Etanol/farmacologia , Artérias Mesentéricas/fisiologia , Resistência Vascular/efeitos dos fármacos , Vasoconstritores/farmacologia , Acetaldeído/farmacologia , Animais , Arginina Vasopressina/farmacologia , Endotelina-1/farmacologia , Etanol/química , Glicerofosfolipídeos/farmacologia , Masculino , Artérias Mesentéricas/química , Artérias Mesentéricas/efeitos dos fármacos , Miografia , Fenilefrina/farmacologia , Cloreto de Potássio/farmacologia , Ratos , Ratos Sprague-Dawley , Vasoconstrição
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