RESUMEN
The hemoparasite Trypanosoma equiperdum belongs to the Trypanozoon subgenus and includes several species that are pathogenic to animals and humans in tropical and subtropical areas across the world. As with all eukaryotic organisms, Ca2+ is essential for these parasites to perform cellular processes thus ensuring their survival across their life cycle. Despite the established paradigm to study proteins related to Ca2+ homeostasis as potential drug targets, so far little is known about Ca2+ entry into trypanosomes. Therefore, in the present study, the presence of a plasma membrane Ca2+-channel in T. equiperdum (TeCC), activated by sphingosine and inhibited by verapamil, is described. The TeCC was cloned and analyzed using bioinformatic resources, which confirmed the presence of several domains, motifs, and a topology similar to the Ca2+ channels found in higher eukaryotes. Biochemical and confocal microscopy assays using antibodies raised against an internal region of human L-type Ca2+ channels indicate the presence of a protein with similar predicted molar mass to the sequence analyzed, located at the plasma membrane of T. equiperdum. Physiological assays based on Fura-2 signals and Mn2+ quenching performed on whole parasites showed a unidirectional Ca2+ entry, which is activated by sphingosine and blocked by verapamil, with the distinctive feature of insensitivity to nifedipine and Bay K 8644. This suggests a second Ca2+ entry for T. equiperdum, different from the store-operated Ca2+ entry (SOCE) previously described. Moreover, the evidence presented here for the TeCC indicates molecular and pharmacological differences with their mammal counterparts, which deserve further studies to evaluate the potential of this channel as a drug target.
Asunto(s)
Esfingosina , Trypanosoma , Animales , Humanos , Esfingosina/farmacología , Verapamilo/farmacología , Membrana Celular/metabolismo , Calcio/metabolismo , MamíferosRESUMEN
BACKGROUND: Hypertrophic and keloid scars are common skin conditions resulting from abnormal wound healing. They can cause itching, pain and have a negative physical and psychological impact on patients' lives. Different approaches are used aiming to improve these scars, including intralesional corticosteroids, surgery and more recently, laser therapy. Since laser therapy is expensive and may have adverse effects, it is critical to evaluate the potential benefits and harms of this therapy for treating hypertrophic and keloid scars. OBJECTIVES: To assess the effects of laser therapy for treating hypertrophic and keloid scars. SEARCH METHODS: In March 2021 we searched the Cochrane Wounds Specialised Register, CENTRAL, MEDLINE, Embase, CINAHL EBSCO Plus and LILACS. To identify additional studies, we also searched clinical trials registries for ongoing and unpublished studies, and scanned reference lists of relevant included studies as well as reviews, meta-analyses, and health technology reports. There were no restrictions with respect to language, date of publication, or study setting. SELECTION CRITERIA: We included randomised controlled trials (RCTs) for treating hypertrophic or keloid scars (or both), comparing laser therapy with placebo, no intervention or another intervention. DATA COLLECTION AND ANALYSIS: Two review authors independently selected studies, extracted the data, assessed the risk of bias of included studies and carried out GRADE assessments to assess the certainty of evidence. A third review author arbitrated if there were disagreements. MAIN RESULTS: We included 15 RCTs, involving 604 participants (children and adults) with study sample sizes ranging from 10 to 120 participants (mean 40.27). Where studies randomised different parts of the same scar, each scar segment was the unit of analysis (906 scar segments). The length of participant follow-up varied from 12 weeks to 12 months. All included trials had a high risk of bias for at least one domain: all studies were deemed at high risk of bias due to lack of blinding of participants and personnel. The variability of intervention types, controls, follow-up periods and limitations with report data meant we pooled data for one comparison (and only two outcomes within this). Several review secondary outcomes - cosmesis, tolerance, preference for different modes of treatment, adherence, and change in quality of life - were not reported in any of the included studies. Laser versus no treatment: We found low-certainty evidence suggesting there may be more hypertrophic and keloid scar improvement (that is scars are less severe) in 585-nm pulsed-dye laser (PDL) -treated scars compared with no treatment (risk ratio (RR) 1.96; 95% confidence interval (CI): 1.11 to 3.45; two studies, 60 scar segments). It is unclear whether non-ablative fractional laser (NAFL) impacts on hypertrophic scar severity when compared with no treatment (very low-certainty evidence). It is unclear whether fractional carbon dioxide (CO2) laser impacts on hypertrophic and keloid scar severity compared with no treatment (very low-certainty evidence). Eight studies reported treatment-related adverse effects but did not provide enough data for further analyses. Laser versus other treatments: We are uncertain whether treatment with 585-nm PDL impacts on hypertrophic and keloid scar severity compared with intralesional corticosteroid triamcinolone acetonide (TAC), intralesional Fluorouracil (5-FU) or combined use of TAC plus 5-FU (very low-certainty evidence). It is also uncertain whether erbium laser impacts on hypertrophic scar severity when compared with TAC (very low-certainty evidence). Other comparisons included 585-nm PDL versus silicone gel sheeting, fractional CO2 laser versus TAC and fractional CO2 laser versus verapamil. However, the authors did not report enough data regarding the severity of scars to compare the interventions. As only very low-certainty evidence is available on treatment-related adverse effects, including pain, charring (skin burning so that the surface becomes blackened), telangiectasia (a condition in which tiny blood vessels cause thread-like red lines on the skin), skin atrophy (skin thinning), purpuric discolorations, hypopigmentation (skin colour becomes lighter), and erosion (loss of part of the top layer of skin, leaving a denuded surface) secondary to blistering, we are not able to draw conclusions as to how these treatments compare. Laser plus other treatment versus other treatment: It is unclear whether 585-nm PDL plus TAC plus 5-FU leads to a higher percentage of good to excellent improvement in hypertrophic and keloid scar severity compared with TAC plus 5-FU, as the certainty of evidence has been assessed as very low. Due to very low-certainty evidence, it is also uncertain whether CO2 laser plus TAC impacts on keloid scar severity compared with cryosurgery plus TAC. The evidence is also very uncertain about the effect of neodymium-doped yttrium aluminium garnet (Nd:YAG) laser plus intralesional corticosteroid diprospan plus 5-FU on scar severity compared with diprospan plus 5-FU and about the effect of helium-neon (He-Ne) laser plus decamethyltetrasiloxane, polydimethylsiloxane and cyclopentasiloxane cream on scar severity compared with decamethyltetrasiloxane, polydimethylsiloxane and cyclopentasiloxane cream. Only very low-certainty evidence is available on treatment-related adverse effects, including pain, atrophy, erythema, telangiectasia, hypopigmentation, regrowth, hyperpigmentation (skin colour becomes darker), and depigmentation (loss of colour from the skin). Therefore, we are not able to draw conclusions as to how these treatments compare. AUTHORS' CONCLUSIONS: There is insufficient evidence to support or refute the effectiveness of laser therapy for treating hypertrophic and keloid scars. The available information is also insufficient to perform a more accurate analysis on treatment-related adverse effects related to laser therapy. Due to the heterogeneity of the studies, conflicting results, study design issues and small sample sizes, further high-quality trials, with validated scales and core outcome sets should be developed. These trials should take into consideration the consumers' opinion and values, the need for long-term follow-up and the necessity of reporting the rate of recurrence of scars to determine whether lasers may achieve superior results when compared with other therapies for treating hypertrophic and keloid scars.
Asunto(s)
Cicatriz Hipertrófica , Hipopigmentación , Queloide , Terapia por Láser , Telangiectasia , Corticoesteroides/uso terapéutico , Adulto , Aluminio , Atrofia , Dióxido de Carbono , Niño , Cicatriz Hipertrófica/etiología , Cicatriz Hipertrófica/radioterapia , Dimetilpolisiloxanos , Erbio , Fluorouracilo , Helio , Humanos , Hipertrofia , Hipopigmentación/etiología , Queloide/etiología , Queloide/radioterapia , Terapia por Láser/efectos adversos , Neodimio , Neón , Dolor/etiología , Geles de Silicona , Telangiectasia/etiología , Triamcinolona Acetonida , Verapamilo , Cicatrización de Heridas , ItrioRESUMEN
BACKGROUND: Idiopathic ventricular tachycardia (VT) in children with structurally normal hearts is generally unrelated to the risk of sudden arrhythmic death. Still, it may be associated with deterioration in the quality of life. VT involving the fascicular conduction system is the most typical form of idiopathic left VT. In this retrospective study, we describe the experience of the clinical presentation, catheter ablation, and long-term follow-up of left fascicular VT in children. METHODS: An electrophysiological study was performed on consecutive children at a single tertiary center. Clinical fascicular left VT was induced by programmed stimulation, and catheter ablation was guided searching for Purkinje potentials. RESULTS: We included 18 patients (0.8 patients/year): 14 (77.8%) males and four females. The mean age of the first VT episode was 8.5 ± 5 years. Intravenous verapamil administration was effective for paroxysmal fascicular VT but not for prevention of recurrences. The mean age at the time of catheter ablation was 11.1 ± 3.8 years (8 months-16 years). The mean weight was 36.8 ± 16.4 kg (8.7-58 kg). A 100% success rate was observed with catheter ablation after repeated procedures without major complications. Mean follow-up was 2.0 ± 1.2 years (1.0-4.0 years, median 1.5), with permanent success in all patients and no antiarrhythmic drug administration. CONCLUSIONS: Fascicular VT has an adverse clinical course in children. In most cases, this condition is drug refractory. Catheter ablation is successful and safe treatment and should represent the first-line approach in symptomatic children.
INTRODUCCIÓN: La taquicardia ventricular (TV) idiopática en niños con corazón estructuralmente normal generalmente no se relaciona con el riesgo de muerte súbita arrítmica, pero puede asociarse con deterioro de la calidad de vida. La TV que involucra el sistema de conducción fascicular es la forma más común de TV izquierda idiopática. En este estudio retrospectivo se describe la experiencia de presentación clínica, ablación con catéter y seguimiento a largo plazo de TV fascicular en niños. MÉTODOS: Se llevó a cabo un estudio electrofisiológico en niños consecutivos en un centro terciario. La TV fascicular clínica se indujo mediante la estimulación programada y la ablación con catéter fue guiada buscando el registro de potenciales de Purkinje. RESULTADOS: Se incluyeron 18 pacientes (0.8 pacientes/año): 14 (77.8%) de sexo masculino y cuatro de sexo femenino. La media de edad a la cual ocurrió el primer episodio fue de 8.5 ± 5 años. La administración intravenosa de verapamilo fue eficaz para la TV fascicular paroxística, pero no para prevención de recurrencias. La media de edad de la ablación con catéter fue de 11.1 ± 3.8 años (8 meses-16 años). La media del peso fue 36.8 ± 16.4 kg (8.7-58 kg). Se observó el 100% de éxito con la ablación con catéter después de procedimientos repetidos sin complicaciones mayores. La media de seguimiento fue de 2.0 ± 1.2 años (1.0-4.0, mediana de 1.5 años) con éxito permanente en todos los pacientes y sin administración de fármacos antiarrítmicos. CONCLUSIONES: En niños, el curso clínico de la TV fascicular es adverso. Además, en la mayoría de los casos, esta condición es refractaria a fármacos. La ablación con catéter resulta exitosa y segura y debe representar el abordaje de primera línea en niños sintomáticos.
Asunto(s)
Ablación por Catéter , Taquicardia Ventricular , Adolescente , Ablación por Catéter/métodos , Niño , Preescolar , Femenino , Humanos , Masculino , Calidad de Vida , Estudios Retrospectivos , Taquicardia Ventricular/diagnóstico , Taquicardia Ventricular/tratamiento farmacológico , Taquicardia Ventricular/cirugía , Verapamilo/uso terapéuticoRESUMEN
INTRODUCTION: Most patients with chronic cardiomyopathy of Chagas disease (CCCD) harbor a secondary cause of coronary microvascular dysfunction (CMD), for which there is no evidence-based therapy. We evaluated the impact of verapamil plus aspirin on symptoms and perfusion abnormalities in patients with CCCD and CMD. METHODS: Consecutive patients with angina pectoris, who had neither coronary artery obstructions nor moderate-severe left ventricular dysfunction (left ventricular ejection fraction > 40%) despite showing wall motion abnormalities on ventriculography, were referred for invasive angiography and tested for Chagas disease. Thirty-two patients with confirmed CCCD and ischemia on stress-rest SPECT myocardial perfusion scintigraphy (MPS) were included. Clinical evaluation, quality of life (EQ-5D/ Seattle Angina Questionnaire), and MPS were assessed before and after 3 months of treatment with oral verapamil plus aspirin (n=26) or placebo (n=6). RESULTS: The mean patient age was 64 years, and 18 (56%) were female. The ischemic index summed difference score (SDS) in MPS was significantly reduced by 55.6% after aspirin+verapamil treatment. A decrease in SDS was observed in 20 (77%) participants, and in 10 participants, no more ischemia could be detected. Enhancements in quality of life were also detected. No change in symptoms or MPS was observed in the placebo group. CONCLUSIONS: This low-cost 3-month treatment for patients diagnosed with CCCD and CMD was safe and resulted in a 55.6% reduction in ischemic burden, symptomatic improvement, and better quality of life.
Asunto(s)
Enfermedad de Chagas , Calidad de Vida , Angina de Pecho/tratamiento farmacológico , Aspirina , Femenino , Humanos , Masculino , Persona de Mediana Edad , Perfusión , Volumen Sistólico , Función Ventricular Izquierda , Verapamilo/uso terapéuticoRESUMEN
AIM: To describe the clinical characteristics and therapeutic options available to paediatric patients with cluster headache. METHOD: Based on a literature search of the medical databases PubMed, LILACS, and Web of Science and using selected descriptors, we carried out a systematic review of case reports on cluster headache in paediatric patients published from 1990 to 2020. RESULTS: Fifty-one patients (29 males, 22 females) with a mean (SD) age of 9 years 7 months (3y 10mo; range 2-16y) were diagnosed with cluster headache. The mean (SD) diagnosis was made 27.8 months (26.2mo) after the onset of cluster headache. Pain occurred at night or on waking up (76.5%) and consisted of 1 to 3 attacks per day (62.7%) lasting 30 to 120 minutes (68.6%). Headaches were unilateral (90.2%), had a pulsatile character (64.7%), and severe intensity (100%). There were autonomic manifestations (90.2%) predominantly ipsilateral to pain, in this order: lacrimation; conjunctival injection; nasal congestion; ptosis; eyelid oedema; and rhinorrhoea. Sumatriptan and oxygen inhalation were the most effective treatments for acute manifestation. Prophylaxis, corticosteroids, verapamil, and gabapentin were the most effective drugs. INTERPRETATION: Due to the small number of published studies, this review could not provide reliable data; however, it appears that cluster headache in children and adolescents is similar to adults, both in clinical characteristics and treatment. What this paper adds Cluster headache in children and adolescents is poorly studied. Cluster headache is uncommon before 10 years of age and diagnosis is difficult in the first few years of life. Treatment of cluster headache in children and adolescents is similar to that used in adults. The notion of the effectiveness of prophylactic treatment is based only on authors' experience.
Asunto(s)
Cefalalgia Histamínica/fisiopatología , Cefalalgia Histamínica/terapia , Adolescente , Corticoesteroides/uso terapéutico , Blefaroptosis/fisiopatología , Niño , Preescolar , Gabapentina/uso terapéutico , Humanos , Terapia por Inhalación de Oxígeno , Rinorrea/fisiopatología , Sumatriptán/uso terapéutico , Lágrimas/fisiología , Vasoconstrictores/uso terapéutico , Verapamilo/uso terapéuticoRESUMEN
Sepsis-induced myocardial dysfunction considerably increases mortality risk in patients with sepsis. Previous studies from our group have shown that sepsis alters the expression of structural proteins in cardiac cells, resulting in cardiomyocyte degeneration and impaired communication between cardiac cells. Caveolin-3 (CAV3) is a structural protein present in caveolae, located in the membrane of cardiac muscle cells, which regulates physiological processes such as calcium homeostasis. In sepsis, there is a disruption of calcium homeostasis, which increases the concentration of intracellular calcium, which can lead to the activation of potent cellular enzymes/proteases which cause severe cellular injury and death. The purpose of the present study was to test the hypotheses that sepsis induces CAV3 overexpression in the heart, and the regulation of L-type calcium channels directly relates to the regulation of CAV3 expression. Severe sepsis increases the expression of CAV3 in the heart, as immunostaining in our study showed CAV3 presence in the cardiomyocyte membrane and cytoplasm, in comparison with our control groups (without sepsis) that showed CAV3 presence predominantly in the plasma membrane. The administration of verapamil, an L-type calcium channel inhibitor, resulted in a decrease in mortality rates of septic mice. This effect was accompanied by a reduction in the expression of CAV3 and attenuation of cardiac lesions in septic mice treated with verapamil. Our results indicate that CAV3 has a vital role in cardiac dysfunction development in sepsis and that the regulation of L-type calcium channels may be related to its expression.
Asunto(s)
Caveolina 3/metabolismo , Corazón/efectos de los fármacos , Sepsis/tratamiento farmacológico , Verapamilo/uso terapéutico , Animales , Canales de Calcio Tipo L , Humanos , Masculino , Ratones , Sepsis/mortalidad , Sepsis/patología , Análisis de Supervivencia , Verapamilo/farmacologíaRESUMEN
In this case report, we describe a 73 year old female with structuraly normal heart that developed shortcoupled torsades de pointes (TdP) resulting in an electrical storm unresponsible to several antiarrhythmic drugs, but fully controlled with verapamil. The critical timing of the ventricular premature beats that initiated TdP corresponded to those that occurred at the peak of the previous T wave. This behavior differentiates this entity from other forms of malignant ventricular arrhythmias in patients with structurally normal heart. It is imperative that the clinical set-up and unique electrocardiographic fingerprint of this unusual malignant entity be assiduously recognized since verapamil can be life-saving in this condition.
Asunto(s)
Torsades de Pointes , Complejos Prematuros Ventriculares , Anciano , Antiarrítmicos/uso terapéutico , Electrocardiografía , Femenino , Humanos , Torsades de Pointes/diagnóstico , Complejos Prematuros Ventriculares/complicaciones , Complejos Prematuros Ventriculares/diagnóstico , Verapamilo/uso terapéuticoRESUMEN
The emergence of resistance requires alternative methods to treat Candida albicans infections. We evaluated efficacy of the efflux pump inhibitor (EPI) verapamil (VER) with fluconazole (FLC) against FLC-resistant (CaR) and -susceptible C. albicans (CaS). The susceptibility of both strains to VER and FLC was determined, as well as the synergism of VER with FLC. Experiments were performed in vitro for planktonic cultures and biofilms and in vivo using Galleria mellonella. Larval survival and fungal recovery were evaluated after treatment with VER and FLC. Data were analyzed by analysis of variance and Kaplan-Meier tests. The combination of VER with FLC at sub-lethal concentrations reduced fungal growth. VER inhibited the efflux of rhodamine 123 and showed synergism with FLC against CaR. For biofilms, FLC and VER alone reduced fungal viability. The combination of VER with FLC at sub-lethal concentrations also reduced biofilm viability. In the in vivo assays, VER and FLC used alone or in combination increased the survival of larvae infected with CaR. Reduction of fungal recovery was observed only for larvae infected with CaR and treated with VER with FLC. VER reverted the FLC-resistance of C. albicans. Based on the results obtained, VER reverted the FLC-resistance of C. albicans and showed synergism with FLC against CaR. VER also increased the survival of G. mellonella infected with CaR and reduced the fungal recovery.
Asunto(s)
Antifúngicos/farmacología , Candida albicans/efectos de los fármacos , Fluconazol/farmacología , Larva/efectos de los fármacos , Proteínas de Transporte de Membrana/metabolismo , Verapamilo/farmacología , Animales , Biopelículas/efectos de los fármacos , Transporte Biológico , Candidiasis/tratamiento farmacológico , Candidiasis/microbiología , Sinergismo Farmacológico , Larva/microbiología , Pruebas de Sensibilidad Microbiana , Mariposas Nocturnas/efectos de los fármacos , Mariposas Nocturnas/microbiologíaRESUMEN
BACKGROUND: Sauromatum guttatum (S. guttatum) is used in the treatment of blood disorders and reportedly has a spasmolytic activity through Ca2+ channel inhibition. OBJECTIVES: The aim of this study was to investigate the antihypertensive potential of S. guttatum in high salt-induced hypertensive Sprague-Dawley (SD) rat model (HSHRs). METHODS: SD rats were divided into normotensive, hypertensive, S. guttatum and verapamil treated groups. S. guttatum crude extract (Sg.Cr) (100, 150 and 300 mg/kg/day) and verapamil (5, 10 and 15 mg/kg/day) were administered orally along with NaCl. Aortic rings and right atrial strips from normotensive rats were used to investigate the underlying mechanisms. The level of statistical significance was set at 5%. RESULTS: Mean arterial pressure decreased in the Sg.Cr and verapamil-treated hypertensive groups in a dose-dependent manner (p < 0.001). In the vascular reactivity study, acetylcholine induced relaxations with an EC50 value of 0.6 µg/mL (0.3-1.0) in Sg.Cr-treated hypertensive rats (300 mg/kg), suggesting endothelial preservation. In isolated normotensive rat aorta, Sg.Cr-treated rats showed vasorelaxation with an EC50 value of 0.15 mg/mL (0.10-0.20), ablated by endothelial denudation or pretreatment with L-NAME and atropine. Sg.Cr treatment caused relaxation against high K+-induced contractions, like verapamil. Sg.Cr showed negative inotropic (82%) and chronotropic effects (56%) in isolated rat atrial preparations reduced with atropine. The phytochemical investigation indicated presence of alkaloids, flavonoids and tannins. CONCLUSION: S. guttatum has a vasodilatory effect through endothelial function preservation, muscarinic receptor-mediated NO release and Ca2+ movement inhibition, while atrial myocardial depressant effect can be linked to the muscarinic receptor. These findings provide pharmacological base for using S. guttatum extract as an antihypertensive medication.
FUNDAMENTO: A Sauromatum guttatum (S. guttatum) é utilizado no tratamento de doenças do sangue e supostamente tem atividade espasmolítica através da inibição dos canais de Ca2+. OBJETIVOS: O objetivo deste estudo foi investigar o potencial anti-hipertensivo de S. guttatum em modelo de rato Sprague-Dawley (SD) com hipertensão induzida por dieta com alto teor de sal (HIDATS). MÉTODOS: Ratos SD foram divididos em normotensos, hipertensos e grupos tratados com verapamil e S. guttatum. Extrato bruto de S. guttatum (Sg.B) (100, 150 e 300 mg/kg/dia) e verapamil (5, 10 e 15 mg/kg/dia) foram administrados por via oral junto com NaCl. Anéis aórticos e faixas do átrio direito de ratos normotensos foram utilizados para investigar os mecanismos subjacentes. O nível de significância estatística adotado foi de 5%. RESULTADOS: A pressão arterial média diminuiu nos grupos hipertensos tratados com Sg.B e verapamil de forma dose-dependente (p <0,001). No estudo de reatividade vascular, a acetilcolina induziu relaxamentos com valor CE50 de 0,6 µg/mL (0,31,0) em ratos hipertensos tratados com Sg.B (300 mg/kg), sugerindo preservação endotelial. Em aorta isolada de rato normotenso, o Sg.B exibiu vasorrelaxamento com valor de CE50 de 0,15 mg/mL (0,10-0,20), após ablação por desnudamento endotelial ou pré-tratamento com L-NAME e atropina. O tratamento com Sg.B causou relaxamento contra contrações induzidas por K+ alto, como o verapamil. O Sg.B mostrou efeitos inotrópicos (82%) e cronotrópicos (56%) negativos em preparações isoladas atriais de ratos reduzidas com atropina. A avaliação fitoquímica indicou a presença de alcaloides, flavonoides e taninos. CONCLUSÃO: O S. guttatum possui efeito vasodilatador através da preservação da função endotelial, liberação de NO mediada pelo receptor muscarínico e inibição do movimento de Ca2+, enquanto o efeito depressor do miocárdio atrial pode estar ligado ao receptor muscarínico. Esses achados fornecem a base farmacológica para o uso do extrato de S. guttatum como um medicamento anti-hipertensivo.
Asunto(s)
Antihipertensivos , Hipertensión , Animales , Antiarrítmicos/uso terapéutico , Antihipertensivos/farmacología , Antihipertensivos/uso terapéutico , Derivados de Atropina/farmacología , Derivados de Atropina/uso terapéutico , Presión Sanguínea , Calcio , Humanos , Hipertensión/tratamiento farmacológico , Extractos Vegetales/farmacología , Ratas , Ratas Sprague-Dawley , Receptores Muscarínicos/uso terapéutico , Vasodilatación , Verapamilo/farmacologíaRESUMEN
Abstract INTRODUCTION: Most patients with chronic cardiomyopathy of Chagas disease (CCCD) harbor a secondary cause of coronary microvascular dysfunction (CMD), for which there is no evidence-based therapy. We evaluated the impact of verapamil plus aspirin on symptoms and perfusion abnormalities in patients with CCCD and CMD. METHODS: Consecutive patients with angina pectoris, who had neither coronary artery obstructions nor moderate-severe left ventricular dysfunction (left ventricular ejection fraction > 40%) despite showing wall motion abnormalities on ventriculography, were referred for invasive angiography and tested for Chagas disease. Thirty-two patients with confirmed CCCD and ischemia on stress-rest SPECT myocardial perfusion scintigraphy (MPS) were included. Clinical evaluation, quality of life (EQ-5D/ Seattle Angina Questionnaire), and MPS were assessed before and after 3 months of treatment with oral verapamil plus aspirin (n=26) or placebo (n=6). RESULTS: The mean patient age was 64 years, and 18 (56%) were female. The ischemic index summed difference score (SDS) in MPS was significantly reduced by 55.6% after aspirin+verapamil treatment. A decrease in SDS was observed in 20 (77%) participants, and in 10 participants, no more ischemia could be detected. Enhancements in quality of life were also detected. No change in symptoms or MPS was observed in the placebo group. CONCLUSIONS: This low-cost 3-month treatment for patients diagnosed with CCCD and CMD was safe and resulted in a 55.6% reduction in ischemic burden, symptomatic improvement, and better quality of life.
Asunto(s)
Humanos , Masculino , Femenino , Calidad de Vida , Enfermedad de Chagas , Perfusión , Volumen Sistólico , Verapamilo/uso terapéutico , Aspirina , Función Ventricular Izquierda , Angina de Pecho/tratamiento farmacológico , Persona de Mediana EdadRESUMEN
Candida is a human fungal pathogen that causes a wide range of diseases. Candida albicans is the main etiologic agent in these diseases; however, infections can be caused by non-albicans Candida species. Virulence factors such as biofilm production, which protect the fungus from host immunity and anti-fungal drugs, are important for the infection. Therefore, available antifungal drugs for candidiasis treatment are limited and the investigation of new and effective drugs is needed. Verapamil is a calcium channel blocker with an inhibitory effect on hyphae development, adhesion, and colonization of C. albicans. In this study, we investigated the effect of verapamil on cell viability and its antifungal and anti-biofilm activity in non-albicans Candida species. Verapamil was not toxic to keratinocyte cells; moreover, C. krusei, C. parapsilosis, and C. glabrata were susceptible to verapamil with a minimal inhibitory concentration (MIC) of 1250 µM; in addition, this drug displayed fungistatic effect at the evaluated concentrations. After treatment with verapamil, reduced viability, biomass, and mitochondrial activity were observed in biofilms of the non-albicans Candida species C. krusei, C. glabrata, and C. parapsilosis. These findings highlight the importance of the study of verapamil as an alternative treatment for infections caused by non-albicans Candida species.
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Antifúngicos , Candida , Antifúngicos/farmacología , Biopelículas , Bloqueadores de los Canales de Calcio/farmacología , Candida albicans , Humanos , Pruebas de Sensibilidad Microbiana , Verapamilo/farmacologíaRESUMEN
INTRODUCTION: Resistance to first-line anti-tuberculosis drugs is a major concern in the treatment of the disease. New strategies, such as the use of efflux pump inhibitors (EPIs), are being investigated to improve the outcome of the treatment. Verapamil (VP), one such inhibitor, was shown to inhibit several efflux pump (EP) Mycobacterium tuberculosis proteins and demonstrate synergic activity with anti-TB drugs.OBJECTIVE: To evaluate the combinatory effect of isoniazid (INH) and VP in M. tuberculosis.METHODS: Minimal inhibitory concentrations and combinatory effects of INH+VP were determined using respectively resazurin microtitre assay plate (REMA) and resazurin drugs combination microtitre assay (REDCA). From the results, we selected three bacilli with different susceptibility profiles and assessed their expression of 10 EP genes through quantitative reverse transcription polymerase chain reaction after exposure to INH, VP and INH + VP for 48 h.RESULTS: A significant reduction of INH MIC was observed in INH-susceptible isolates upon combination with VP. In brief, gene expression assays revealed expression patterns that could be correlated with each resistance profile, presence or absence of gene mutations and combinatory effect with VP.CONCLUSION: Combining VP with INH showed important results in drug-susceptible strains, and clinical trials on combined VP + anti-TB drugs should be discussed.
Asunto(s)
Mycobacterium tuberculosis , Tuberculosis Resistente a Múltiples Medicamentos , Antituberculosos/farmacología , Proteínas Bacterianas/genética , Expresión Génica , Humanos , Isoniazida/farmacología , Proteínas de Transporte de Membrana/genética , Pruebas de Sensibilidad Microbiana , Mutación , Mycobacterium tuberculosis/genética , Tuberculosis Resistente a Múltiples Medicamentos/tratamiento farmacológico , Verapamilo/farmacologíaRESUMEN
AIMS: Increased activity of calpain-1 and matrix metalloproteinase (MMP)-2 was observed in different models of arterial hypertension and contribute to thicken the left ventricle (LV) walls and to hypertrophy cardiac myocytes. MMP-2 activity may be regulated by calpain-1 via bioactive molecules activation such as transforming growth factor (TGF)-ß in cardiovascular diseases. This study analyzed whether calpain-1 causes cardiac hypertrophy and dysfunction by modulating the expression and activity of MMP-2 in renovascular hypertension. MAIN METHODS: Male Wistar rats were submitted to two kidneys, one clip (2K1C) model of hypertension or sham surgery and were treated with verapamil (VRP, 8 mg/kg/bid) by gavage from the second to tenth week post-surgery. Systolic blood pressure (SBP) was weekly assessed by tail-cuff plethysmography and morphological and functional parameters of LV were analyzed by echocardiography. MMP-2 activity was analyzed by in situ and gelatin zymography, while calpain-1 activity by caseinolytic assay. MMP-2, calpain-1, TGF-ß and MMP-14/TIMP-2 levels were identified in the LV by western blots. Fluorescence assays were performed to evaluate oxidative stress, MMP-2 and calpain-1 levels. KEY FINDINGS: SBP increased in 2K1C rats and was unaltered by VRP. However, VRP notably ameliorated hypertension-induced increase in LV thickness. VRP decreased hypertension-induced enhances in calpain-1 and MMP-2 activities, oxidative stress and mature TGF-ß levels. Treatment with VRP also decreased the accentuated MMP-14/TIMP-2 levels in 2K1C. SIGNIFICANCE: Treatment with VRP decreases calpain-1 and MMP-2 activities and also reduces TGF-ß and MMP-14/TIMP-2 levels in the LV of hypertensive rats, thus contributing to ameliorate cardiac hypertrophy.
Asunto(s)
Calpaína/metabolismo , Cardiomegalia/tratamiento farmacológico , Regulación de la Expresión Génica/efectos de los fármacos , Hipertensión/complicaciones , Metaloproteinasa 2 de la Matriz/metabolismo , Remodelación Ventricular/efectos de los fármacos , Verapamilo/farmacología , Animales , Calpaína/genética , Cardiomegalia/etiología , Masculino , Metaloproteinasa 2 de la Matriz/genética , Ratas , Ratas Wistar , Vasodilatadores/farmacologíaAsunto(s)
Enfermedad de la Arteria Coronaria , Hipertensión , Presión Sanguínea , Femenino , Humanos , Indoles , VerapamiloRESUMEN
O guia Q8(R2) do guia ICH descreve Qualidade por Design (QbD) como "uma abordagem sistemática para desenvolvimento farmacêutico que começa com objetivos predefinidos e enfatiza produto, entendimento e controle dos processos, baseado em dados científicos sólidos e gestão do risco da qualidade". Os métodos analíticos são considerados parte integrante do desenvolvimento farmacêutico. Assim, a Qualidade por Design Analítico (AQbD) é justificável e recomendada para obter flexibilidade regulatória, reduzir os resultados fora de especificação, obter um alto grau de robustez e um método analítico econômico. O Planejamento de Experimentos (DoE) é um conjunto de ferramentas estatísticas que inclui delineamentos de triagem e otimização, no qual os fatores são sistematicamente variados para determinar seus efeitos nas respostas, o que permite a determinação de quais fatores são os mais significantes, a identificação de qual configuração de fatores resulta em respostas otimizadas e a identificação de interações entre os fatores. As abordagens QbD e AQbD permitem a melhoria contínua ao longo do ciclo de vida do produto farmacêutico e do método analítico, inclusive para reduzir a variabilidade do produto, melhorar o desempenho do processo, reduzir resultados fora da especificação, melhorar o desempenho analítico, entre outros. O Cloridrato de Verapamil foi escolhido como molécula teste para desenvolvimento do projeto. Na primeira etapa do estudo foi realizado uma triagem com 13 fatores e 20 experimentos, utilizando o delineamento Plackett-Burman, seguiu-se para a próxima etapa com 7 fatores e 16 experimentos através do delineamento fatorial fracionado (Res.: IV). A etapa de otimização foi realizada com 3 fatores e 20 experimentos utilizando o delineamento central composto. Após todas as etapas do estudo, as seguintes condições foram consideradas ideais: Fase móvel A - Tampão formiato de amônio 10 mM pH 3,0, Fase móvel B - Amoníaco 2,0% em acetonitrila, eluição do tipo gradiente, coluna cromatográfica XSelect CSH C18 (100mm x 4,6mm x 3,5 µm), fluxo de 0,7 mL/min, volume de injeção de 2 µL para teor e 10 µL para produtos de degradação. Os métodos desenvolvidos são robustos, validados e indicativos de estabilidade
The ICH guide Q8 (R2) describes Quality by Design as "a systematic approach to pharmaceutical development that begins with predefined goals and emphasizes product, understanding and control of processes, based on solid scientific data and Quality Risk Management ". Analytical methods are considered an integral part of pharmaceutical development. Thus, the application of QbD approach to analytical method development is justifiable and a recommended strategy to attain regulatory flexibility, to reduce out-of-specification results, to achieve a high degree of robustness and a cost-effective analytical method. DoE is a set of statistical tools which include screening designs and optimization designs. In DoE approach, the controlled input factors are systematically varied to determine their effects on the output responses, which allows the determination of the most important input factors, the identification of input factors setting leading to optimized output responses, and the identification of interactions between input factors. The QbD and AQbD approach allows the continuous improvement throughout the lifecycle of pharmaceutical product and analytical method, including to reduce product variability, to improve process performance, to reduce out-of-specification results, to improve analytical performance, among others. Verapamil Hydrochloride was chosen as a test molecule for the development of the project. In the first phase of the study, a 13-factor and 20-experiment screening was performed using the Plackett-Burman design, followed by the 7-factor and 16-experiment next stage through fractional factorial design (Res .: IV). The optimization step was performed with 3 factors and 20 experiments using the composite central design. After performing all the study steps, the following conditions were considered ideal: Mobile Phase A - 10 mM ammonium formate buffer pH 3.0, Mobile Phase B - 2.0% ammonia in acetonitrile, gradient elution, column chromatographic XSelect CSH C18 (100mm x 4.6mm x 3.5µm), flow rate of 0.7ml / min, injection volume of 2µL for assay and 10µL for degradation products. The methods developed are robust, validated and stability indicating
Asunto(s)
Métodos de Análisis de Laboratorio y de Campo/métodos , Dibujo , Métodos , Acetonitrilos/efectos adversos , Preparaciones Farmacéuticas , Verapamilo , Tamizaje Masivo , Desarrollo de Medicamentos/instrumentaciónRESUMEN
The membrane-based efflux pump systems are recognized to have an important role in pathogenicity and drug resistance in Mycobacterium tuberculosis by the extrusion of toxic substrates and drugs from the inner bacillus. This study aimed to investigate the in vitro interaction of Verapamil (VP), an efflux pump inhibitor, with the classical first-line anti-tuberculosis drug isoniazid (INH) in resistant and susceptible M. tuberculosis clinical isolates. Seven multidrug-resistant (MDR), three INH monoresistant and four susceptible M. tuberculosis clinical isolates were tested for the INH and VP combination by modified Resazurin Microtiter Assay Plate (REMA). Fractional Inhibitory Concentration (FIC) and Modulation Factor (MF) were determined. The INH plus VP combination showed no significant change in the Minimum inhibitory concentration (MIC) values of INH (FIC≥ 0.5; MF=1 or 2).The use of VP in tuberculosis therapy should be managed carefully, considering the resistance caused by specific mutation in katG and inhA genes, in which the use of these EPIs may have no success. The use of EPIs as an adjunctive drug in the anti-tuberculosis therapy should be further investigated on a larger number of M. tuberculosis clinical isolates with different resistant profile.
Asunto(s)
Verapamilo/antagonistas & inhibidores , Mycobacterium tuberculosis/aislamiento & purificación , Antituberculosos , Bacillus/clasificación , Tuberculosis/patología , Técnicas In Vitro/métodos , Resistencia a Medicamentos , Preparaciones Farmacéuticas/análisis , Pruebas de Sensibilidad Microbiana/instrumentación , Isoniazida/agonistasAsunto(s)
Induración Peniana/terapia , Guías de Práctica Clínica como Asunto , Administración Tópica , Antiinflamatorios no Esteroideos/uso terapéutico , Terapia Combinada , Tratamiento con Ondas de Choque Extracorpóreas/métodos , Humanos , Interferón alfa-2/uso terapéutico , Iontoforesis/métodos , Masculino , Colagenasa Microbiana/uso terapéutico , Vasodilatadores/uso terapéutico , Verapamilo/uso terapéutico , Vitamina E/uso terapéuticoRESUMEN
BACKGROUND: Colonic vascular lesion secondary to verapamil overdose is mediated by free radicals, forming vascular microtrombos and endotoxin generation, being a difficult diagnosis. CLINICAL CASE: A 27-year-old female is admitted with an acute abdomen of 4 days after an event referred for a suicidal attempt due to an overdose of verapamil, operating surgically where there is a right transmural colon necrosis, performing a right hemicolectomy with terminal ileostomy. CONCLUSIONS: Recognize and properly treat an acute abdomen, not always reach an adequate diagnosis, so a thorough history could conclude.
ANTECEDENTES: La lesión vascular colónica secundaria a la sobredosis de verapamilo, es mediada por radicales libres, formando microtrombos vasculares y generación de endotoxinas, siendo un diagnostico difícil. CASO CLÍNICO: Femenino de 27 años, ingresa con abdomen agudo de 4 días posteriores a un evento remitido de intento suicida por sobredosis de verapamilo, interviniéndose quirúrgicamente donde se halla necrosis colónica transmural derecha, realizando hemicolectomía derecha con ileostomía terminal. CONCLUSIONES: Reconocer y tratar de forma adecuada un abdomen agudo, no siempre se suele llegar a un adecuado diagnostico, por lo cual una minuciosa anamnesis lograría concluirlo.
Asunto(s)
Abdomen Agudo/cirugía , Colectomía , Colon/patología , Ileostomía , Isquemia/inducido químicamente , Circulación Esplácnica/efectos de los fármacos , Abdomen Agudo/inducido químicamente , Absceso Abdominal/etiología , Adulto , Colon/irrigación sanguínea , Drenaje , Femenino , Humanos , Hipotensión/inducido químicamente , Obstrucción Intestinal/etiología , Obstrucción Intestinal/cirugía , Isquemia/patología , Necrosis , Intento de Suicidio , Taquicardia/inducido químicamente , Verapamilo/envenenamientoRESUMEN
AIM: To evaluate modulatory effect of verapamil (VP) in rifampicin (RIF) activity and its effect in efflux pumps (EPs) transcript levels in Mycobacterium tuberculosis. MATERIALS & METHODS: RIF and VP minimal inhibitory concentration, combinatory effect and detection of mutations were determined in 16 isolates. EPs transcript levels were determined in four isolates by real-time PCR after exposure to drugs. RESULTS: VP showed good combinatory effect among RIF-resistant isolates. This effect was also observed in the relative transcript levels of EPs, mainly after 72 h of exposure, depending on the EP gene, genotype and the resistance profile of the isolate. CONCLUSION: Additional regulatory mechanisms in the EP activities, as well as, interactions with other drug-specific resistance mechanisms need further investigation in M. tuberculosis.
Asunto(s)
Antituberculosos/farmacología , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Mycobacterium tuberculosis/efectos de los fármacos , Rifampin/farmacología , Verapamilo/farmacología , Proteínas Bacterianas/efectos de los fármacos , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Brasil , Catalasa/genética , ARN Polimerasas Dirigidas por ADN/genética , Sinergismo Farmacológico , Quimioterapia Combinada , Regulación Bacteriana de la Expresión Génica , Genotipo , Humanos , Proteínas de Transporte de Membrana/efectos de los fármacos , Proteínas de Transporte de Membrana/genética , Proteínas de Transporte de Membrana/metabolismo , Pruebas de Sensibilidad Microbiana , Mutación , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/metabolismo , Oxidorreductasas/genética , Factores de Tiempo , Tuberculosis Resistente a Múltiples Medicamentos/microbiologíaRESUMEN
Fungal resistance is the major problem related to fluconazole treatments. This study aims to develop innovative lipid core nanocapsules and nanostructured lipid carriers containing fluconazole, to study in vitro antifungal activity and to assess the possibility of resistance reversion in Candida albicans, C. glabrata, C. krusei, and C. tropicalis isolates. The action mechanism of nanoparticles was investigated through efflux pumps and scanning electron microscopy studies. The lipid core nanocapsules and nanostructured lipid carriers were prepared by interfacial deposition of preformed polymer and high-pressure homogenization methods, respectively. Both nanostructures presented sizes below 250 nm, SPAN < 1.6, negative zeta potential, pH slightly acid, high drug content and controlled drug release. The nanostructured lipid carriers were unable to reverse the fungal resistance. Lipid core nanoparticles displayed advantages such as a reduction in the effective dose of fluconazole and resistance reversion in all isolates tested - with multiple mechanisms of resistance. The main role of the supramolecular structure and the composition of the nanoparticles on antifungal mechanisms of action were discussed. The results achieved through this study have an impact on clinical therapy, with a potential application in the treatment of fungal infections caused by resistant isolates of Candida spp.