RESUMEN

BACKGROUND: The presence of ovarian-type stroma defines mucinous cystic neoplasm (MCN). Criteria for surgical resection differ between current consensus guidelines (IAP, AGA, and Europe). This meta-analysis aims to describe pre-surgical clinical parameters that predict malignant transformation of MCN of the pancreas. METHODS: A systematic review and meta-analysis of articles published from 2006 to the time of manuscript authorship in December 2022. The electronic databases included English publications in Ovid MEDLINE In-Process & Other Non-Indexed Citations, Ovid MEDLINE, Ovid EMBASE, and Scopus. RESULTS: 17 studies were identified and included 1058 patients with MCN treated with pancreatectomy. The mean cohort age was 48.2 years (standard deviation [SD] ± 7.9) with an expected female predominance (96 %). The presenting symptom for most was abdominal pain (55.6 %), however, nearly 20 % of patients were asymptomatic. Most patients were treated with distal pancreatectomy (70.5 %), and the mean tumor size was 45 mm. The rate of invasive cancer was 13.8 %. Cysts with mural nodules had a higher risk of developing invasive tumors than those that did not (OR 26.47, 95%CI 12.57-55.74, p < 0.001, I2:0 %). Other clinical factors such as the presence of intramural calcifications or an elevated serum CA 19-9 (>37U/mL) were not predictive of malignancy. CONCLUSION: The present meta-analysis did not clarify establishing reliable predictors for malignant transformation other than mural modularity, which may represent tumors that have already undergone transformation. It may be used as a criterion in treatment decision-making.

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BACKGROUND: Stroke is a leading cause of death worldwide, with oxidative stress and calcium overload playing significant roles in the pathophysiology of the disease. Ozone, renowned for its potent antioxidant properties, is commonly employed as an adjuvant therapy in clinical settings. Nevertheless, it remains unclear whether ozone therapy on parthanatos in cerebral ischemia-reperfusion injury (CIRI). This study aims to investigate the impact of ozone therapy on reducing parthanatos during CIRI and to elucidate the underlying mechanism. METHODS: Hydrogen peroxide (H2O2) was utilized to mimic the generation of reactive oxygen species (ROS) in SH-SY5Y cell reperfusion injury in vitro, and an in vivo ischemic stroke model was established. Ozone saline was introduced for co-culture or intravenously administered to mice. Apoptosis and oxidative stress were assessed using flow cytometry and immunofluorescence. Western blotting was utilized to examine the expression of parthanatos signature proteins. The mechanism by which ozone inhibits parthanatos was elucidated through inhibiting PPARg or Nrf2 activity. RESULTS: The findings demonstrated that ozone mitigated H2O2-induced parthanatos by either upregulating nuclear factor erythroid 2-related factor 2 (Nrf2) or activating peroxisome proliferator-activated receptorg (PPARg). Furthermore, through the use of calcium chelators and ROS inhibitors, it was discovered that ROS directly induced parthanatos and facilitated intracellular calcium elevation. Notably, a malignant feedback loop between ROS and calcium was identified, further amplifying the induction of parthanatos. Ozone therapy exhibited its efficacy by increasing PPARg activity or enhancing the Nrf2 translation, thereby inhibiting ROS production induced by H2O2. Concurrently, our study demonstrated that ozone treatment markedly inhibited parthanatos in stroke-afflicted mice. Additionally, ozone therapy demonstrated significant neuroprotective effects on cortical neurons, effectively suppressing parthanatos. CONCLUSIONS: These findings contribute valuable insights into the potential of ozone therapy as a therapeutic strategy for reducing parthanatos during CIRI, highlighting its impact on key molecular pathways associated with oxidative stress and calcium regulation.

Asunto(s)

Modelos Animales de Enfermedad , Accidente Cerebrovascular Isquémico , Estrés Oxidativo , Ozono , Especies Reactivas de Oxígeno , Ozono/farmacología , Ozono/uso terapéutico , Animales , Accidente Cerebrovascular Isquémico/tratamiento farmacológico , Ratones , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Daño por Reperfusión , Masculino , Peróxido de Hidrógeno/metabolismo , Humanos , Factor 2 Relacionado con NF-E2/metabolismo , Apoptosis/efectos de los fármacos , Ratones Endogámicos C57BL , Calcio/metabolismo

RESUMEN

OBJECTIVE: In the context of postoperative anal pain, understanding the intricate mechanisms and effective interventions is paramount. This study investigates the role of Muscarinic Acetylcholine Receptors (mAChRs) and the IP3-Ca2+-CaM signaling pathway in a rat model of postoperative anal pain, exploring the potential analgesic effects of electroacupuncture. METHODS: Comprehensive approaches involving mechanical sensitivity assays, Western blotting, immunohistochemistry, and intracellular calcium concentration measurement were used. RESULTS: The authors found elevated mAChRs expression in the postoperative pain model. Antagonizing mAChRs reduced pain sensitivity and attenuated the IP3-Ca2+-CaM pathway. Remarkably, electroacupuncture treatment further mitigated pain, potentially by suppressing this signaling cascade. INTERPRETATION: These findings reveal a novel connection between mAChRs and the IP3-Ca2+-CaM pathway in postoperative anal pain and suggest electroacupuncture as a promising avenue for pain relief through these mechanisms, offering insights into innovative strategies for postoperative pain management.

Asunto(s)

Electroacupuntura , Hemorreoidectomía , Dolor Postoperatorio , Ratas Sprague-Dawley , Receptores Muscarínicos , Transducción de Señal , Animales , Electroacupuntura/métodos , Dolor Postoperatorio/terapia , Masculino , Hemorreoidectomía/métodos , Receptores Muscarínicos/metabolismo , Puntos de Acupuntura , Canal Anal/cirugía , Modelos Animales de Enfermedad , Western Blotting , Ratas , Inmunohistoquímica , Calcio/metabolismo , Resultado del Tratamiento

RESUMEN

Objetivo: Describir las características clínicas de las pacientes con cáncer de ovario epitelial en el Instituto Oncológico Nacional de Panamá́. Métodos: Se realizó un estudio descriptivo en el Instituto Oncológico Nacional. Se revisaron todos los expedientes de las pacientes con cáncer de ovario epitelial desde enero 2012 a diciembre 2021. Aquellas con diagnóstico histopatológico confirmado de cáncer de ovario epitelial primario fueron incluidas en el estudio. Entre las variables evaluadas, se encontraban edad, lugar de procedencia, grupo histológico, etapa al diagnóstico, tratamiento y recidivas. Resultados: Se incluyeron 547 pacientes. El 49% (266) de las pacientes residían en la provincia de Panamá. La edad media fue de 56 años (DE:13.2). El 71% (386) estaban aseguradas. El 80% (439) presentaron síntomas relacionados al cáncer. El 80% (442) presentaron valores de CA-125 elevados. El 59% (321) eran de tipo seroso. El 62% (338) fueron tratadas con una combinación de cirugía y quimioterapia. En etapa I, el 19% (27) recayeron mientras que en etapa III, el 39% (103). El 80% (138) de las recidivas recibieron sólo quimioterapia. Conclusiones: En general, el cáncer epitelial se presentó en etapas avanzadas en mujeres mayores de 40 años asociado a sintomatología y valores de CA-125 elevados. El tipo histológico más frecuente fue el seroso y el tratamiento habitual fue una combinación de cirugía y quimioterapia. El porcentaje de muerte fue directamente proporcional con la etapa al diagnóstico. (provisto por Infomedic International)

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Objetivo: Describir las características clínicas de las pacientes con cáncer de ovario epitelial en el Instituto Oncológico Nacional de Panamá́. Métodos: Se realizó un estudio descriptivo en el Instituto Oncológico Nacional. Se revisaron todos los expedientes de las pacientes con cáncer de ovario epitelial desde enero 2012 a diciembre 2021. Aquellas con diagnóstico histopatológico confirmado de cáncer de ovario epitelial primario fueron incluidas en el estudio. Entre las variables evaluadas, se encontraban edad, lugar de procedencia, grupo histológico, etapa al diagnóstico, tratamiento y recidivas. Resultados: Se incluyeron 547 pacientes. El 49% (266) de las pacientes residían en la provincia de Panamá. La edad media fue de 56 años (DE:13.2). El 71% (386) estaban aseguradas. El 80% (439) presentaron síntomas relacionados al cáncer. El 80% (442) presentaron valores de CA-125 elevados. El 59% (321) eran de tipo seroso. El 62% (338) fueron tratadas con una combinación de cirugía y quimioterapia. En etapa I, el 19% (27) recayeron mientras que en etapa III, el 39% (103). El 80% (138) de las recidivas recibieron sólo quimioterapia. Conclusiones: En general, el cáncer epitelial se presentó en etapas avanzadas en mujeres mayores de 40 años asociado a sintomatología y valores de CA-125 elevados. El tipo histológico más frecuente fue el seroso y el tratamiento habitual fue una combinación de cirugía y quimioterapia. El porcentaje de muerte fue directamente proporcional con la etapa al diagnóstico. (provided by Infomedic International)

RESUMEN

This study aimed to elucidate vincristine (VCR)-induced peripheral neuropathy in aged rats, a poorly understood neurotoxicity. Both young and old Wistar rats were administered VCR (0.1 mg/kg, intraperitoneally (i.p.)) and compared to age-matched controls (0.9% saline; 10 mg/mL, i.p.). Mechanical (MN) and thermal nociceptive (TN) responses were assessed on days 0, 6, 11, and 17. Locomotor response, cognitive ability, and anxious-like behavior were evaluated on days 14, 15, and 16. Results showed MN and TN responses in both young and old VCR-exposed rats. In old rats, VCR exacerbated MN (on days 6, 11, and 17) and TN (on days 6 and 17) responses. VCR also induced cognitive impairments and anxiety-like behavior. Histological analysis revealed Wallerian degeneration in the spinal cords of VCR-exposed rats accompanied by macrophage migration. Furthermore, VCR increased Ca2+-ATPase activity while inhibiting Na+, K+-ATPase activity in young and old rats. VCR altered the homeostasis of Mg2+-ATPase activity. Lipid peroxidation and nitrite and nitrate levels increased in young and old rats exposed to VCR. This study provides valuable insights into VCR's mechanistic pathways in aged rats, emphasizing the need for further research in this area.

Asunto(s)

Adenosina Trifosfatasas , Envejecimiento , Estrés Oxidativo , Enfermedades del Sistema Nervioso Periférico , Ratas Wistar , Vincristina , Degeneración Walleriana , Animales , Vincristina/farmacología , Vincristina/toxicidad , Enfermedades del Sistema Nervioso Periférico/inducido químicamente , Enfermedades del Sistema Nervioso Periférico/patología , Enfermedades del Sistema Nervioso Periférico/metabolismo , Estrés Oxidativo/efectos de los fármacos , Ratas , Degeneración Walleriana/inducido químicamente , Degeneración Walleriana/patología , Degeneración Walleriana/metabolismo , Masculino , Envejecimiento/efectos de los fármacos , Adenosina Trifosfatasas/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Antineoplásicos Fitogénicos/farmacología , Médula Espinal/efectos de los fármacos , Médula Espinal/metabolismo , Médula Espinal/patología , ATPasa Intercambiadora de Sodio-Potasio/metabolismo

RESUMEN

BACKGROUND: While calcium is known to play a crucial role in mammalian sperm physiology, how it flows in and out of the male gamete is not completely understood. Herein, we investigated the involvement of Na+/Ca2+ exchangers (NCX) in mammalian sperm capacitation. Using the pig as an animal model, we first confirmed the presence of NCX1 and NCX2 isoforms in the sperm midpiece. Next, we partially or totally blocked Ca2+ outflux (forward transport) via NCX1/NCX2 with different concentrations of SEA0400 (2-[4-[(2,5-difluorophenyl)methoxy]phenoxy]-5-ethoxyaniline; 0, 0.5, 5 and 50 µM) and Ca2+ influx (reverse transport) with SN6 (ethyl 2-[[4-[(4-nitrophenyl)methoxy]phenyl]methyl]-1,3-thiazolidine-4-carboxylate; 0, 0.3, 3 or 30 µM). Sperm were incubated under capacitating conditions for 180 min; after 120 min, progesterone was added to induce the acrosome reaction. At 0, 60, 120, 130, and 180 min, sperm motility, membrane lipid disorder, acrosome integrity, mitochondrial membrane potential (MMP), tyrosine phosphorylation of sperm proteins, and intracellular levels of Ca2+, reactive oxygen species (ROS) and superoxides were evaluated. RESULTS: Partial and complete blockage of Ca2+ outflux and influx via NCX induced a significant reduction of sperm motility after progesterone addition. Early alterations on sperm kinematics were also observed, the effects being more obvious in totally blocked than in partially blocked samples. Decreased sperm motility and kinematics were related to both defective tyrosine phosphorylation and mitochondrial activity, the latter being associated to diminished MMP and ROS levels. As NCX blockage did not affect the lipid disorder of plasma membrane, the impaired acrosome integrity could result from reduced tyrosine phosphorylation. CONCLUSIONS: Inhibition of outflux and influx of Ca2+ triggered similar effects, thus indicating that both forward and reverse Ca2+ transport through NCX exchangers are essential for sperm capacitation.

Asunto(s)

Calcio , Intercambiador de Sodio-Calcio , Capacitación Espermática , Animales , Masculino , Capacitación Espermática/efectos de los fármacos , Intercambiador de Sodio-Calcio/metabolismo , Intercambiador de Sodio-Calcio/efectos de los fármacos , Calcio/metabolismo , Porcinos , Espermatozoides/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Motilidad Espermática/efectos de los fármacos , Reacción Acrosómica/efectos de los fármacos , Potencial de la Membrana Mitocondrial/efectos de los fármacos

RESUMEN

Wnt signaling plays an essential role in cellular processes like development, maturation, and function maintenance. Xenopus laevis oocytes are a suitable model to study not only the development but also the function of different receptors expressed in their membranes, like those receptors expressed in the central nervous system (CNS) including Frizzled 7. Here, using frog oocytes and recordings of endogenous membrane currents in a two-electrode path configuration along with morphological observations, we evaluated the role of the non-canonical Wnt-5a ligand in oocytes. We found that acute application of Wnt-5a generated changes in endogenous calcium-dependent currents, entry oscillatory current, the membrane's outward current, and induced membrane depolarization. The incubation of oocytes with Wnt-5a caused a reduction of the membrane potential, potassium outward current, and protected the ATP current in the epithelium/theca removed (ETR) model. The oocytes exposed to Wnt-5a showed increased viability and an increase in the percentage of the germinal vesicle breakdown (GVBD), at a higher level than the control with progesterone. Altogether, our results suggest that Wnt-5a modulates different aspects of oocyte structure and generates calcium-dependent endogenous current alteration and GVDB process with a change in membrane potential at different concentrations and times of the exposition. These results help to understand the cellular effect of Wnt-5a and present the use of Xenopus oocytes to explore the mechanism that could impact the activation of Wnt signaling.

RESUMEN

In the last two decades, several working groups in the international psychoanalytic community have been interested in the development of systematic tools for psychodynamic diagnosis, case formulation and treatment planning. Such psychodynamic diagnostic manuals are efforts to systematically integrate an enormous and rich amount of historically partialized and dispersed information, but which constitute the substantial contribution of psychoanalysis to the field of mental health. The aim of the present article is to provide an updated review on this kind of systematic tools for diagnosis, case formulation and therapeutic planning, designed for the field of psychodynamic approaches. To this end, we describe the aims and structure of: 1) the Psychodynamic Diagnostic Manual 2 (PDM-2), 2) the Operationalized Psychodynamic Diagnosis (OPD-2/OPD-3) and 3) the Operationalized Psychodynamic Diagnosis for Children and adolescents 2 (OPD-CA-2). The contributions of these current tools to clinical practice and empirical research are discussed, as well as the need to disseminate these types of instruments in our regional context.

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En las últimas dos décadas, diversos grupos de trabajo de la comunidad psicoanalítica internacional se han interesado por el desarrollo de herramientas sistemáticas para el diagnóstico, la formulación de los casos y la planificación del tratamiento psicodinámico. Este tipo de manuales diagnósticos psicodinámicos son esfuerzos de integración sistemática de una enorme y rica cantidad de información históricamente parcializada y dispersa, pero que constituye el aporte sustancial del psicoanálisis al campo de la salud mental. El objetivo del presente artículo es ofrecer una revisión actualizada sobre esta clase de herramientas sistemáticas de diagnóstico, formulación del caso y planificación terapéutica, diseñadas para el campo de los abordajes psicodinámicos. A estos fines, se describe la estructura y los objetivos de: 1) el Manual Diagnóstico Psicodinámico 2 (PDM-2), 2) el Diagnóstico Psicodinámico Operacionalizado (OPD-2/OPD-3) y 3) el Diagnóstico Psicodinámico Operacionalizado Infanto-Juvenil 2 (OPD-IJ-2).Se discuten las contribuciones de estas herramientas actuales para la práctica clínica y la investigación empírica, así como la necesidad de difundir este tipo de instrumentos en nuestro contexto regional.

Asunto(s)

Trastornos Mentales , Psicoterapia Psicodinámica , Humanos , Psicoterapia Psicodinámica/métodos , Trastornos Mentales/terapia , Trastornos Mentales/diagnóstico

RESUMEN

BACKGROUND: Exposure of humans and animals to heavy metals is increasing day-by-day; thus, lead even today remains of significant public health concern. According to CDC, blood lead reference value (BLRV) ranges from 3.5 µg/dl to 5 µg/dl in adults. Recently, almost 2.6% decline in male fertility per year has been reported but the cause is not well established. Lead (Pb2+) affects the size of testis, semen quality, and secretory functions of prostate. But the molecular mechanism(s) of lead toxicity in sperm cells is not clear. Thus, present study was undertaken to evaluate the adverse effects of lead acetate at environmentally relevant exposure levels (0.5, 5, 10 and 20 ppm) on functional and molecular dynamics of spermatozoa of bucks following in vitro exposure for 15 min and 3 h. RESULTS: Lead significantly decreased motility, viable count, and motion kinematic patterns of spermatozoa like curvilinear velocity, straight-line velocity, average path velocity, beat cross frequency and maximum amplitude of head lateral displacement even at 5 ppm concentration. Pb2+ modulated intracellular cAMP and Ca2+ levels in sperm cells through L-type calcium channels and induced spontaneous or premature acrosome reaction (AR) by increasing tyrosine phosphorylation of sperm proteins and downregulated mitochondrial transmembrane potential. Lead significantly increased DNA damage and apoptosis as well. Electron microscopy studies revealed Pb2+ -induced deleterious effects on plasma membrane of head and acrosome including collapsed cristae in mitochondria. CONCLUSIONS: Pb2+ not only mimics Ca2+ but also affects cellular targets involved in generation of cAMP, mitochondrial transmembrane potential, and ionic exchange. Lead seems to interact with Ca2+ channels because of charge similarity and probably enters the sperm cell through these channels and results in hyperpolarization. Our findings also indicate lead-induced TP and intracellular Ca2+ release in spermatozoa which in turn may be responsible for premature acrosome exocytosis which is essential feature of capacitation for fertilization. Thus, lead seems to reduce the fertilizing capacity of spermatozoa even at 0.5 ppm concentrations.

Asunto(s)

Reacción Acrosómica , Acrosoma , Calcio , Plomo , Motilidad Espermática , Espermatozoides , Masculino , Espermatozoides/efectos de los fármacos , Calcio/metabolismo , Motilidad Espermática/efectos de los fármacos , Animales , Acrosoma/efectos de los fármacos , Plomo/toxicidad , Reacción Acrosómica/efectos de los fármacos , AMP Cíclico/metabolismo , Bovinos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Análisis de Semen , Daño del ADN/efectos de los fármacos , Compuestos Organometálicos/toxicidad , Compuestos Organometálicos/farmacología

RESUMEN

To meet the current demand for lead-free piezoelectric ceramics, a novel sol-gel synthesis route is presented for the preparation of Ba0.85Ca0.15Ti0.9Zr0.1O3 doped with cerium (Ce = 0, 0.01, and 0.02 mol%) and vanadium (V = 0, 0.3, and 0.4 mol%). X-ray diffraction patterns reveal the formation of a perovskite phase (space group P4mm) for all samples after calcination at 800 °C and sintering at 1250, 1350, and 1450 °C, where it is proposed that both dopants occupy the B site. Sintering studies show that V doping allows the sintering temperature to be reduced to at least 1250 °C. Undoped BCZT samples sintered at the same temperature show reduced functional properties compared to V-doped samples, i.e., d33 values increase by an order of magnitude with doping. The dissipation factor tan δ decreases with increasing sintering temperature for all doping concentrations, while the Curie temperature TC increases for all V-doped samples, reaching 120 °C for high-concentration co-doped samples. All results indicate that vanadium doping can facilitate the processing of BCZT at lower sintering temperatures without compromising performance while promoting thermal property stability.

RESUMEN

Chagas disease vectors can ingest several times their own volume in blood with each meal. This ad libitum feeding causes an intense process of diuresis, inducing the insect to eliminate a large quantity of urine and faeces. To ensure diuresis, the speed of circulation of the haemolymph is increased. The Triatominae circulatory system is quite simple, including the dorsal vessel, which pumps haemolymph in an anterograde direction. The return is caused by peristaltic contractions of the anterior midgut. Triatominae insects can spend several weeks without feeding, meaning that most of the time, the insect is in a resting condition. Although the mechanisms controlling the circulation of the haemolymph during post-prandial diuresis have been largely analysed, the mechanisms controlling it during resting conditions are poorly understood. In this study, we analysed several canonical pathways (i.e. L-type VGCC, GPCR, RyR, IP3R) and a novel system represented by the recently characterized Piezo proteins. Our results show that during the resting condition, haemolymph circulation depends on a cross-talk between myogenic activity, inhibitory and stimulatory cellular messengers, and Piezo proteins. This report also unveils for the first time the existence of a putative Piezo protein in Hemiptera.

Asunto(s)

Hemolinfa , Rhodnius , Animales , Rhodnius/fisiología , Proteínas de Insectos/metabolismo , Insectos Vectores/fisiología , Enfermedad de Chagas/transmisión , Descanso/fisiología

RESUMEN

OBJECTIVE: To demonstrate a high-yield molecular diagnostic workflow for lateralized overgrowth (LO), a congenital condition with abnormal enlargement of body parts, and to classify it by molecular genetics. STUDY DESIGN: We categorized 186 retrospective cases of LO diagnosed between 2003 and 2023 into suspected Beckwith-Wiedemann spectrum, PIK3CA-related overgrowth spectrum (PROS), vascular overgrowth, or isolated LO, based on initial clinical assessments, to determine the appropriate first-tier molecular tests and tissue for analysis. Patients underwent testing for 11p15 epigenetic abnormalities or somatic variants in genes related to PI3K/AKT/mTOR, vascular proliferation, and RAS-MAPK cascades using blood or skin DNA. For cases with negative initial tests, a sequential cascade molecular approach was employed to improve diagnostic yield. RESULTS: This approach led to a molecular diagnosis in 54% of cases, 89% of cases consistent with initial clinical suspicions, and 11% reclassified. Beckwith-Wiedemann spectrum was the most common cause, with 43% of cases exhibiting 11p15 abnormalities. PIK3CA-related overgrowth spectrum had the highest confirmation rate, with 74% of clinically diagnosed patients showing a PIK3CA variant. Vascular overgrowth demonstrated significant clinical overlap with other syndromes. A molecular diagnosis of isolated LO proved challenging, with only 21% of cases classifiable into a specific condition. CONCLUSIONS: LO is underdiagnosed from a molecular viewpoint and to date has had no diagnostic guidelines, which is crucial for addressing potential cancer predisposition, enabling precision medicine treatments, and guiding management. This study sheds light on the molecular etiology of LO, highlighting the importance of a tailored diagnostic approach and of selecting appropriate testing to achieve the highest diagnostic yield.

Asunto(s)

Síndrome de Beckwith-Wiedemann , Fosfatidilinositol 3-Quinasa Clase I , Humanos , Estudios Retrospectivos , Femenino , Masculino , Fosfatidilinositol 3-Quinasa Clase I/genética , Niño , Preescolar , Síndrome de Beckwith-Wiedemann/genética , Síndrome de Beckwith-Wiedemann/diagnóstico , Lactante , Trastornos del Crecimiento/diagnóstico , Trastornos del Crecimiento/genética , Adolescente

RESUMEN

Pannexin1 hemichannels (Panx1 HCs) are found in the membrane of most mammalian cells and communicate the intracellular and extracellular spaces, enabling the passive transfer of ions and small molecules. They are involved in physiological and pathophysiological conditions. During apoptosis, the C-terminal tail of Panx1 is proteolytically cleaved, but the permeability features of hemichannels and their role in cell death remain elusive. To address these topics, HeLa cells transfected with full-length human Panx1 (fl-hPanx1) or C-terminal truncated hPanx1 (Δ371hPanx1) were exposed to alkaline extracellular saline solution, increasing the activity of Panx1 HCs. The Δ371hPanx1 HC was permeable to DAPI and Etd+, but not to propidium iodide, whereas fl-hPanx1 HC was only permeable to DAPI. Furthermore, the cytoplasmic Ca2+ signal increased only in Δ371hPanx1 cells, which was supported by bioinformatics approaches. The influx of Ca2+ through Δ371hPanx1 HCs was necessary to promote cell death up to about 95% of cells, whereas the exposure to alkaline saline solution without Ca2+ failed to induce cell death, and the Ca2+ ionophore A23187 promoted more than 80% cell death even in fl-hPanx1 transfectants. Moreover, cell death was prevented with carbenoxolone or 10Panx1 in Δ371hPanx1 cells, whereas it was undetectable in HeLa Panx1-/- cells. Pretreatment with Ferrostatin-1 and necrostatin-1 did not prevent cell death, suggesting that ferroptosis or necroptosis was not involved. In comparison, zVAD-FMK, a pancaspase inhibitor, reduced death by ~60%, suggesting the involvement of apoptosis. Therefore, alkaline pH increases the activity of Δ371hPanx1HCs, leading to a critical intracellular free-Ca2+ overload that promotes cell death.

Asunto(s)

Calcio , Conexinas , Proteínas del Tejido Nervioso , Humanos , Conexinas/metabolismo , Conexinas/genética , Células HeLa , Calcio/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Proteínas del Tejido Nervioso/genética , Apoptosis , Muerte Celular , Señalización del Calcio

RESUMEN

Our research aimed to elucidate the mechanism by which aurintricarboxylic acid (ATA) inhibits plasma membrane Ca2+-ATPase (PMCA), a crucial enzyme responsible for calcium transport. Given the pivotal role of PMCA in cellular calcium homeostasis, understanding how it is inhibited by ATA holds significant implications for potentially regulating physiopathological cellular processes in which this pump is involved. Our experimental findings revealed that ATA employs multiple modes of action to inhibit PMCA activity, which are influenced by ATP but also by the presence of calcium and magnesium ions. Specifically, magnesium appears to enhance this inhibitory effect. Our experimental and in-silico results suggest that, unlike those reported in other proteins, ATA complexed with magnesium (ATA·Mg) is the molecule that inhibits PMCA. In summary, our study presents a novel perspective and establishes a solid foundation for future research efforts aimed at the development of new pharmacological molecules both for PMCA and other proteins.

Asunto(s)

Ácido Aurintricarboxílico , Calcio , Magnesio , ATPasas Transportadoras de Calcio de la Membrana Plasmática , Magnesio/metabolismo , Magnesio/farmacología , Ácido Aurintricarboxílico/farmacología , ATPasas Transportadoras de Calcio de la Membrana Plasmática/metabolismo , ATPasas Transportadoras de Calcio de la Membrana Plasmática/antagonistas & inhibidores , Calcio/metabolismo , Adenosina Trifosfato/metabolismo , Membrana Celular/metabolismo , Membrana Celular/efectos de los fármacos , Animales , Humanos

RESUMEN

Lithium therapy received approval during the 1970s, and it has been used for its antidepressant, antimanic, and anti-suicidal effects for acute and long-term prophylaxis and treatment of bipolar disorder (BPD). These properties have been well established; however, the molecular and cellular mechanisms remain controversial. In the past few years, many studies demonstrated that at the cellular level, lithium acts as a regulator of neurogenesis, aging, and Ca2+ homeostasis. At the molecular level, lithium modulates aging by inhibiting glycogen synthase kinase-3ß (GSK-3ß), and the phosphatidylinositol (PI) cycle; latter, lithium specifically inhibits inositol production, acting as a non-competitive inhibitor of inositol monophosphatase (IMPase). Mitochondria and peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) have been related to lithium activity, and its regulation is mediated by GSK-3ß degradation and inhibition. Lithium also impacts Ca2+ homeostasis in the mitochondria modulating the function of the lithium-permeable mitochondrial Na+-Ca2+exchanger (NCLX), affecting Ca2+ efflux from the mitochondrial matrix to the endoplasmic reticulum (ER). A close relationship between the protease Omi, GSK-3ß, and PGC-1α has also been established. The purpose of this review is to summarize some of the intracellular mechanisms related to lithium activity and how, through them, neuronal aging could be controlled.

Asunto(s)

Senescencia Celular , Compuestos de Litio , Neuronas , Neuronas/efectos de los fármacos , Compuestos de Litio/farmacología , Fármacos Neuroprotectores/farmacología , Enzimas/metabolismo , Inositol/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Calcio/metabolismo , Humanos , Animales , Senescencia Celular/efectos de los fármacos

RESUMEN

Background: Sarcomeric hypertrophic cardiomyopathy (HCM) must be differentiated from phenotypically similar conditions because clinical management and prognosis may greatly differ. Patients with unexplained left ventricular hypertrophy require an early, confirmed genetic diagnosis through diagnostic or predictive genetic testing. We tested the feasibility and practicality of the application of a 17-gene next-generation sequencing (NGS) panel to detect the most common genetic causes of HCM and HCM phenocopies, including treatable phenocopies, and report detection rates. Identification of transthyretin cardiac amyloidosis (ATTR-CA) and Fabry disease (FD) is essential because of the availability of disease-specific therapy. Early initiation of these treatments may lead to better clinical outcomes. Methods: In this international, multicenter, cross-sectional pilot study, peripheral dried blood spot samples from patients of cardiology clinics with an unexplained increased left ventricular wall thickness (LVWT) of ≥13 mm in one or more left ventricular myocardial segments (measured by imaging methods) were analyzed at a central laboratory. NGS included the detection of known splice regions and flanking regions of 17 genes using the Illumina NextSeq 500 and NovaSeq 6000 sequencing systems. Results: Samples for NGS screening were collected between May 2019 and October 2020 at cardiology clinics in Colombia, Brazil, Mexico, Turkey, Israel, and Saudi Arabia. Out of 535 samples, 128 (23.9%) samples tested positive for pathogenic/likely pathogenic genetic variants associated with HCM or HCM phenocopies with double pathogenic/likely pathogenic variants detected in four samples. Among the 132 (24.7%) detected variants, 115 (21.5%) variants were associated with HCM and 17 (3.2%) variants with HCM phenocopies. Variants in MYH7 (n=60, 11.2%) and MYBPC3 (n=41, 7.7%) were the most common HCM variants. The HCM phenocopy variants included variants in the TTR (n=7, 1.3%) and GLA (n=2, 0.4%) genes. The mean (standard deviation) ages of patients with HCM or HCM phenocopy variants, including TTR and GLA variants, were 42.8 (17.9), 54.6 (17.0), and 69.0 (1.4) years, respectively. Conclusions: The overall diagnostic yield of 24.7% indicates that the screening strategy effectively identified the most common forms of HCM and HCM phenocopies among geographically dispersed patients. The results underscore the importance of including ATTR-CA (TTR variants) and FD (GLA variants), which are treatable disorders, in the differential diagnosis of patients with increased LVWT of unknown etiology.

RESUMEN

L-type calcium channels are essential for the excitation-contraction coupling in cardiac muscle. The CaV1.2 channel is the most predominant isoform in the ventricle which consists of a multi-subunit membrane complex that includes the CaV1.2 pore-forming subunit and auxiliary subunits like CaVα2δ and CaVß2b. The CaV1.2 channel's C-terminus undergoes proteolytic cleavage, and the distal C-terminal domain (DCtermD) associates with the channel core through two domains known as proximal and distal C-terminal regulatory domain (PCRD and DCRD, respectively). The interaction between the DCtermD and the remaining C-terminus reduces the channel activity and modifies voltage- and calcium-dependent inactivation mechanisms, leading to an autoinhibitory effect. In this study, we investigate how the interaction between DCRD and PCRD affects the inactivation processes and CaV1.2 activity. We expressed a 14-amino acid peptide miming the DCRD-PCRD interaction sequence in both heterologous systems and cardiomyocytes. Our results show that overexpression of this small peptide can displace the DCtermD and replicate the effects of the entire DCtermD on voltage-dependent inactivation and channel inhibition. However, the effect on calcium-dependent inactivation requires the full DCtermD and is prevented by overexpression of calmodulin. In conclusion, our results suggest that the interaction between DCRD and PCRD is sufficient to bring about the current inhibition and alter the voltage-dependent inactivation, possibly in an allosteric manner. Additionally, our data suggest that the DCtermD competitively modifies the calcium-dependent mechanism. The identified peptide sequence provides a valuable tool for further dissecting the molecular mechanisms that regulate L-type calcium channels' basal activity in cardiomyocytes.

Asunto(s)

Canales de Calcio Tipo L , Miocitos Cardíacos , Canales de Calcio Tipo L/metabolismo , Canales de Calcio Tipo L/genética , Canales de Calcio Tipo L/química , Animales , Miocitos Cardíacos/metabolismo , Humanos , Células HEK293 , Ratas , Dominios Proteicos

RESUMEN

The Cupressaceae family includes species considered to be medicinal. Their essential oil is used for headaches, colds, cough, and bronchitis. Cedar trees like Chamaecyparis lawsoniana (C. lawsoniana) are commonly found in urban areas. We investigated whether C. lawsoniana exerts some of its effects by modifying airway smooth muscle (ASM) contractility. The leaves of C. lawsoniana (363 g) were pulverized mechanically, and extracts were obtained by successive maceration 1:10 (w:w) with methanol/CHCl3. Guinea pig tracheal rings were contracted with KCl, tetraethylammonium (TEA), histamine (HIS), or carbachol (Cch) in organ baths. In the Cch experiments, tissues were pre-incubated with D-600, an antagonist of L-type voltage-dependent Ca2+ channels (L-VDCC) before the addition of C. lawsoniana. Interestingly, at different concentrations, C. lawsoniana diminished the tracheal contractions induced by KCl, TEA, HIS, and Cch. In ASM cells, C. lawsoniana significantly diminished L-type Ca2+ currents. ASM cells stimulated with Cch produced a transient Ca2+ peak followed by a sustained plateau maintained by L-VDCC and store-operated Ca2+ channels (SOCC). C. lawsoniana almost abolished this last response. These results show that C. lawsoniana, and its active metabolite quercetin, relax the ASM by inhibiting the L-VDCC and SOCC; further studies must be performed to obtain the complete set of metabolites of the extract and study at length their pharmacological properties.

Asunto(s)

Calcio , Chamaecyparis , Contracción Muscular , Músculo Liso , Extractos Vegetales , Quercetina , Tráquea , Animales , Cobayas , Músculo Liso/efectos de los fármacos , Músculo Liso/metabolismo , Contracción Muscular/efectos de los fármacos , Quercetina/farmacología , Quercetina/química , Tráquea/efectos de los fármacos , Tráquea/metabolismo , Extractos Vegetales/farmacología , Extractos Vegetales/química , Chamaecyparis/química , Calcio/metabolismo , Masculino , Bloqueadores de los Canales de Calcio/farmacología , Histamina/metabolismo , Canales de Calcio Tipo L/metabolismo , Hojas de la Planta/química

RESUMEN

OBJECTIVE: This study aimed to investigate the effect of Esketamine (ESK) on the Hypoxia/Reoxygenation (H/R) injury of cardiomyocytes by regulating TRPV1 and inhibiting the concentration of intracellular Ca2+. METHODS: The H/R injury model of H9c2 cardiomyocytes was established after 4h hypoxia and 6h reoxygenation. H9c2 cells were treated with different concentrations of ESK or TRPV1 agonist capsaicin (10 µM) or TRPV1 inhibitor capsazepine (1 µM). Cell viability was detected by CCK-8 method, and apoptosis by flow cytometry. Intracellular Ca2+ concentration was evaluated by Fluo-4 AM. LDH, MDA, SOD, and GSH-Px were detected with corresponding commercial kits. TRPV1 and p-TRPV1 proteins were detected by Western blot. RESULTS: After H/R, H9c2 cell viability decreased, apoptosis increased, intracellular Ca2+ concentration increased, LDH and MDA levels increased, SOD and GSH-Px levels decreased, and p-TRPV1 expression increased. ESK treatment rescued these changes induced by H/R. After up-regulating TRPV1, the protective effect of ESK on H/R injury of H9c2 cells was weakened, while down-regulating TRPV1 could further protect against H/R injury. CONCLUSION: ESK alleviates H/R injury of cardiomyocytes by regulating TRPV1 expression and inhibiting intracellular Ca2+ concentration.

Asunto(s)

Apoptosis , Calcio , Capsaicina/análogos & derivados , Supervivencia Celular , Ketamina , Miocitos Cardíacos , Canales Catiónicos TRPV , Canales Catiónicos TRPV/metabolismo , Canales Catiónicos TRPV/efectos de los fármacos , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Calcio/metabolismo , Supervivencia Celular/efectos de los fármacos , Apoptosis/efectos de los fármacos , Animales , Ketamina/farmacología , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Ratas , Capsaicina/farmacología , Hipoxia de la Célula/efectos de los fármacos , Línea Celular , Citometría de Flujo , Estrés Oxidativo/efectos de los fármacos , Western Blotting

RESUMEN

Chronic kidney disease (CKD) is a prevalent health concern associated with various pathological conditions, including hypertensive nephropathy. Mesangial cells are crucial in maintaining glomerular function, yet their involvement in CKD pathogenesis remains poorly understood. Recent evidence indicates that overactivation of Pannexin-1 (Panx1) channels could contribute to the pathogenesis and progression of various diseases. Although Panx1 is expressed in the kidney, its contribution to the dysfunction of renal cells during pathological conditions remains to be elucidated. This study aimed to investigate the impact of Panx1 channels on mesangial cell function in the context of hypertensive nephropathy. Using an Ang II-infused mouse model and primary mesangial cell cultures, we demonstrated that in vivo exposure to Ang II sensitizes cultured mesangial cells to show increased alterations when they are subjected to subsequent in vitro exposure to Ang II. Particularly, mesangial cell cultures treated with Ang II showed elevated activity of Panx1 channels and increased release of ATP. The latter was associated with enhanced basal intracellular Ca2+ ([Ca2+]i) and increased ATP-mediated [Ca2+]i responses. These effects were accompanied by increased lipid peroxidation and reduced cell viability. Crucially, all the adverse impacts evoked by Ang II were prevented by the blockade of Panx1 channels, underscoring their critical role in mediating cellular dysfunction in mesangial cells. By elucidating the mechanisms by which Ang II negatively impacts mesangial cell function, this study provides valuable insights into the pathogenesis of renal damage in hypertensive nephropathy.