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RESUMEN Antecedentes: la cirugía hepática ha evolucionado con el correr del tiempo. No existe bibliografía publicada sobre experiencia previa de hepatectomía en Corrientes. Nuestro grupo de trabajo se desenvuelve en el ámbito público y privado. Objetivo: describir los resultados iniciales de una serie consecutiva de pacientes con hepatectomías de distinta extensión, por el mismo equipo quirúrgico. Material y métodos: trabajo retrospectivo, observacional, descriptivo, sobre datos de las historias clínicas y libros de quirófano de pacientes con hepatectomías realizadas entre septiembre de 2019 y enero de 2023, en la práctica pública y privada. Resultados: fueron intervenidos 27 pacientes, con media de edad 53 años (25-72); 16 eran mujeres. El abordaje fue convencional en 25 casos y laparoscópico en 2. Se realizaron 4 hepatectomías mayores y 23 menores. Los diagnósticos fueron de patología maligna en 22 oportunidades y benigna en 5. La sobrevida a los 90 días fue de 96,2%. La mortalidad fue de 1 paciente (3,7%). Con respecto a las complicaciones, 2 pacientes (7,4%) presentaron abscesos hepáticos en el posoperatorio, 2 pacientes (7,4%) requirieron reintervención quirúrgica por sangrado, con buena evolución posterior y alta hospitalaria. Conclusión: la morbimortalidad en la serie descripta estuvo en relación con lo comunicado por otros autores.
ABSTRACT Background: Liver surgery has evolved over time. There are no prior publications on the experience of liver surgery in the province of Corrientes. Our work group operates in both the public and private sectors. Objective: The aim of this study was to describe the initial results of a consecutive series of patients undergoing different types of liver resections, performed by the same surgical team. Material and methods: We conducted a retrospective, observational, and descriptive study based on data from the medical records and operating room records of patients undergoing liver resection at public and private institutions from September 2019 to January 2023. Results: A total of 27 patients were operated on; mean age was 53 years (25-72) and 16 were women. We used the conventional approach in 25 cases and laparoscopy in 2. Four procedures were major liver resections and 23 were minor liver resections, The diagnoses were cancer in 22 cases and benign conditions in 5. Survival at 90 days was 96.2% One patient died (3.7%). The complications included postoperative liver abscesses in 2 patients (7.4%) and re-operation due to bleeding in 2 patients (7.4%), who had a subsequent favorable course and were discharged from the hospital. Conclusion: The morbidity and mortality in the described series were similar to those reported by other authors.
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The abbreviated measurement of coping strategies is useful for monitoring and identifying the effects of stress. The Coping strategy indicator-Short version (CSI-S, including the dimensions of seeking support, problem solving and avoidance strategies) is a new adaptation of the full version of this indicator, and additional evidence of its validity is needed. Psychology students (n = 125) from a public university in Lima, Peru, were recruited to help provide such evidence of validity in terms of internal structure, reliability and associations with other variables (perceived stress and general efficacy in cope with difficulties), which were evaluated using nonparametric item response theory procedures. Support-seeking and problem-solving items from the Mokken scale and the avoidance scale exhibited limitations. The correlations between the scales were moderate or low and exhibited theoretical consistency, and the relationship with perceived stress highlighted the predictive capacity of avoidance and problem-solving strategies. In general, the CSI-S exhibits suitable psychometric properties; however, the avoidance score requires further examination or reconstruction of its items.
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Capacidades de Enfrentamento , Estudantes , Humanos , Reprodutibilidade dos Testes , Psicometria , PeruRESUMO
The D1R and D3R receptors functionally and synergistically interact in striatonigral neurons. Dopaminergic denervation turns this interaction antagonistic, which is correlated with a decrement in D3nf isoform and an increment in D3R membranal expression. The mechanisms of such changes in D3R are attributed to the dysregulation of the expression of their isoforms. The cause and mechanism of this phenomenon remain unknown. Dopaminergic denervation produces a decrement in D1R and PKA activity; we propose that the lack of phosphorylation of PTB (regulator of alternative splicing) by PKA produces the dysregulation of D3R splicing and changes D3R functionality. By using in silico analysis, we found that D3R mRNA has motifs for PTB binding and, by RIP, co-precipitates with PTB. Moreover, D1R activation via PKA promotes PTB phosphorylation. Acute and 5-day D1R blockade decreases the expression of D3nf mRNA. The 5-day treatment reduces D3R, D3nf, and PTB protein in the cytoplasm and increases D3R in the membrane and PTB in the nucleus. Finally, the blockade of D1R mimics the effect of dopaminergic denervation in D1R and D3R signaling. Thus, our data indicate that through PKAâPTB, D1R modulates D3R splicing, expression, and signaling, which are altered during D1R blockade or the lack of stimulation in dopaminergic denervation.
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Growth Arrest-Specific 1 (Gas1) is a pleiotropic protein with different functions, in the adult kidney Gas1 acts as an endogenous inhibitor of cell proliferation but it is also necessary for the maintenance and proliferation of Renal Progenitor Cells (RPC) during early development, thus it fulfills important functions in the adult kidney. However, it is not known whether or not Gas1 is expressed during postnatal development, a critical stage for renal maturation. For this reason, the main objective of this work was to characterize the expression pattern of Gas1 in the different regions of the kidney by immunofluorescence and Western blot analysis during the postnatal development of the rat. We found that Gas1 is present and has a differential expression pattern in the various regions of the nephron during postnatal development. We observed that the highest levels of expression of Gas1 occur in the adult, however, Gas1 is also expressed in RPC and interestingly, the expression of RPC markers such as the Neural cell adhesion molecule (NCAM) and Cluster of differentiation 24 (CD24) were found to have an inverse pattern of expression to Gas1 (decreases as the kidney matures) during postnatal renal maturation, this indicates a role for Gas1 in the regulation of renal cell proliferation at this stage of development.
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Proteínas de Ciclo Celular , Néfrons , Ratos , Animais , Proteínas de Ciclo Celular/metabolismo , Proliferação de Células , Néfrons/metabolismo , Células-Tronco/metabolismo , Células Epiteliais/metabolismo , Proteínas Ligadas por GPI/metabolismoRESUMO
Glioblastoma (GBM) is a very aggressive tumor that presents vascularization, necrosis and is resistant to chemotherapy and radiotherapy. Current treatments are not effective eradicating GBM, thus, there is an urgent need to develop novel therapeutic strategies against GBM. AZD5363, AZD8542, curcumin and resveratrol, are widely studied for the treatment of cancer and in the present study we explored the effects of the administration of combined treatments with AZD5363, AZD8542, curcumin or resveratrol on human GBM cells. We found that the combined treatments with AZD5363+AZD8542+Curcumin and AZD8542+Curcumin+Resveratrol inhibit the PI3K/AKT and SHH survival pathways by decreasing the activity of AKT, the reduction of the expression of SMO, pP70S6k, pS6k, GLI1, p21 and p27, and the activation of caspase-3 as a marker of apoptosis. These results provide evidence that the combined treatments AZD5363+AZD8542+Curcumin and AZD8542+Curcumin+Resveratrol have the potential to be an interesting option against GBM.
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The cellular prion protein (PrPC) is a metal-binding biomolecule that can interact with different protein partners involved in pivotal physiological processes, such as neurogenesis and neuronal plasticity. Recent studies profile copper and PrPC as important players in the pathological mechanisms of Alzheimer's disease and cancer. Although the copper-PrPC interaction has been characterized extensively, the role of the metal ion in the physiological and pathological roles of PrPC has been barely explored. In this article, we discuss how copper binding and proteolytic processing may impact the ability of PrPC to recruit protein partners for its functional roles. The importance to dissect the role of copper-PrPC interactions in health and disease is also underscored.
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Doença de Alzheimer , Neoplasias , Química Bioinorgânica , Cobre/metabolismo , Humanos , Proteínas Priônicas/químicaRESUMO
Glioblastoma (GB) is the most common and aggressive primary brain tumour in adult humans. Therapeutic resistance and tumour recurrence after surgical removal contribute to poor prognosis for glioblastoma patients. Men are known to be more likely than women to develop an aggressive form of GB, and differences in sex steroids have emerged as a leading explanation for this finding. Studies indicate that the metabolism and proliferation of GB-derived cells are increased by sex steroids, the expression of androgen receptors (ARs) and the synthesis of androgens and oestrogens, suggesting that these hormones have a role in the tumour pathogenesis. The expression of aromatase, the enzyme that converts androgens to oestrogens, has been reported in glial cells and GB cell lines. Thus, it was necessary to test whether the steroidogenic enzymes involved in androgen synthesis are expressed in GB cells. Therefore, here, we investigated the expression of four key enzymes involved in androgen synthesis in human-derived GB cells. U87 cells were cultured in Dulbecco's modified Eagle medium plus foetal bovine serum and antibiotics on slides for immunocytochemistry or immunofluorescence. U87, LN229 and C6 cells were also cultured in multi-well chambers to obtain proteins for Western blotting. We used primary antibodies against 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17α-hydroxilase/17,20-lyase (P450c17), 17ß-hydroxysteroid dehydrogenase (17ß-HSD) and 5α-reductase. Immunocytochemistry, and immunofluorescence results revealed that glioblastoma cells express 3ß-HSD, P450c17, 17ß-HSD and 5α-reductase proteins in their cytoplasm. Moreover, Western blot analyses revealed bands corresponding to the molecular weight of these four enzymes in the three GB cell lines. Thus, glioblastoma cells have the key enzymatic machinery necessary to synthesize androgens, and these enzymes might be useful targets for new therapeutic approaches.
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Androgênios , Glioblastoma , 17-Hidroxiesteroide Desidrogenases/metabolismo , Adulto , Androgênios/metabolismo , Colestenona 5 alfa-Redutase , Feminino , Humanos , Masculino , Oxirredutases , Esteroide 17-alfa-Hidroxilase/metabolismoRESUMO
Huntington's disease (HD) is an inherited neurodegenerative disorder caused by an abnormal CAG repeat expansion in the huntingtin gene coding for a protein with an elongated polyglutamine sequence. HD patients present choreiform movements, which are caused by the loss of neurons in the striatum and cerebral cortex. Previous reports indicate that the absence of the aryl hydrocarbon receptor (AhR) protects mice from excitotoxic insults and increases the transcription of neurotrophic factors. Based on these data, we evaluated the effects of the lack of the AhR on a mice model of HD, generating a double transgenic mouse, expressing human mutated huntingtin (R6/1 mice) and knockout for the AhR. Our results show that the body weight of 30-week-old double transgenic mice is similar to that of R6/1 mice; however, feet clasping, an indicative of neuronal damage in the R6/1 animals, was not observed. In addition, motor coordination and ambulatory behavior in double transgenic mice did not deteriorate over time as occur in the R6/1 mice. Moreover, the anxiety behavior of double transgenic mice was similar to wild type mice. Interestingly, astrogliosis is also reduced in the double transgenic mice. The present data demonstrate that the complete loss of the AhR reduces the motor and behavioral deterioration observed in R6/1 mice, suggesting that the pharmacological modulation of the AhR could be a therapeutic target in HD.
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Comportamento Animal/fisiologia , Gliose/fisiopatologia , Proteína Huntingtina/genética , Doença de Huntington/metabolismo , Doença de Huntington/fisiopatologia , Atividade Motora/fisiologia , Receptores de Hidrocarboneto Arílico/fisiologia , Animais , Modelos Animais de Doenças , Doença de Huntington/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , FenótipoRESUMO
BACKGROUND: Glioblastomas (GBMs) are the main primary brain tumors in adults with almost 100% recurrence rate. Patients with lateral ventricle proximal GBMs (LV-GBMs) exhibit worse survival compared to distal locations for unknown reasons. One hypothesis is the proximity of these tumors to the cerebrospinal fluid (CSF) and its chemical cues that can regulate cellular phenotype. We therefore investigated the role of CSF on GBM gene expression and the role of a CSF-induced gene, SERPINA3, in GBM malignancy in vitro and in vivo. METHODS: We utilized human CSF and GBM brain tumor-initiating cells (BTICs). We determined the impact of SERPINA3 expression in glioma patients using The Cancer Genome Atlas (TCGA) database. SERPINA3 expression changes were evaluated at mRNA and protein levels. The effects of knockdown (KD) and overexpression (OE) of SERPINA3 on cell migration, viability and cell proliferation were evaluated. Stem cell characteristics on KD cells were evaluated by differentiation and colony formation experiments. Tumor growth was studied by intracranial and flank injections. RESULTS: GBM-CSF increased BTIC migration accompanied by upregulation of the SERPINA3 gene. In patient samples and TCGA data, we observed SERPINA3 to correlate directly with brain tumor grade and indirectly with GBM patient survival. SERPINA3 KD induced a decrease in cell proliferation, migration, invasion, and stem cell characteristics, while SERPINA3 OE increased cell migration. In vivo, SERPINA3 KD BTICs showed increased survival in a murine model. CONCLUSIONS: SERPINA3 plays a key role in GBM malignancy and its inhibition results in a better outcome using GBM preclinical models.
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Neoplasias Encefálicas , Glioblastoma , Células-Tronco Neoplásicas , alfa 1-Antiquimotripsina , Adulto , Animais , Neoplasias Encefálicas/genética , Linhagem Celular Tumoral , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Glioblastoma/genética , Humanos , Camundongos , SerpinasRESUMO
RESUMEN Antecedentes: la lesión quirúrgica de la vía biliar representa un gran problema de salud y puede surgir ante cualquier cirujano que realice una colecistectomía. Objetivos: el objetivo del trabajo fue presentar nuestra experiencia en reparación de la vía biliar, ana lizando la morbimortalidad y la incidencia de dicha patología en nuestro Servicio. Material y métodos: estudio retrospectivo descriptivo; se tomaron las variables de las historias clínicas de los pacientes en un período de 8 años, de enero de 2011 a julio de 2019 donde fueron admitidos 19 pacientes que presentaron lesión quirúrgica de la vía biliar en el Hospital José Ramón Vidal de la provincia de Corrientes, Argentina. Resultados: 12 pacientes fueron tratados quirúrgicamente mediante hepático-yeyuno anastomosis, 2 por bihepático-yeyuno anastomosis y dos mediante sutura término-terminal bilio-biliar sobre tubo de Kehr. Tres pacientes fueron tratados mediante colocación de stent y dilatación posterior mediante colangiopancreatografia retrógrada endoscópica. Conclusión: los cirujanos deben entrenarse para disminuir al mínimo la posibilidad de una lesión. El objetivo de una colecistectomía debería ser no lesionar la vía biliar.
ABSTRACT Background: Bile duct injury represents a serious health problem and can occur after any cholecystectomy. Objectives: The aim of this study was to report our experience in repairing bile duct injuries analyzing morbidity, mortality and its incidence in our department. Material and Methods: We conducted a retrospective and descriptive study. The information was retrieved form the medical records of 19 patients with bile duct injury hospitalized at the Hospital José Ramón Vidal, Corrientes, Argentina, between January 2011 and July 2019. Results: A Roux-en-Y hepaticojejunostomy was performed in 12 patients, double hepaticojejunostomy in two patients, and two patients were treated with end-to-end ductal anastomosis with suture over a T tube. Three patients underwent endoscopic retrograde cholangiopancreatography with stent placement and dilation. Conclusion: Surgeons should be trained to avoid the possibility of bile duct injury. The main goal of cholecystectomy should be to avoid this complication.
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Humanos , Masculino , Feminino , Adolescente , Adulto , Pessoa de Meia-Idade , Idoso , Adulto Jovem , Ferimentos e Lesões/cirurgia , Ductos Biliares/lesões , Colecistectomia Laparoscópica/efeitos adversos , Argentina , Ductos Biliares/cirurgia , Anastomose Cirúrgica , Colangiografia , Indicadores de Morbimortalidade , Epidemiologia Descritiva , Estudos Retrospectivos , Angiografia por Tomografia Computadorizada , Hospitais PúblicosRESUMO
INTRODUCTION: Pancreatic carcinoma cells exhibit a pronounced tendency to invade along and through intra and extrapancreatic nerves, even during the early stages of the disease, a phenomenon called perineural invasion (PNI). Thus, we sought to determine the effects of the simultaneous expression of soluble forms of GAS1 and PTEN (tGAS1 and PTEN-L) inhibiting tumor growth and invasiveness. MATERIALS AND METHODS: We employed a lentiviral system to simultaneously express tGAS1 and PTEN-L; in order to determine the effects of the treatments, cell viability and apoptosis as well as the expression of the transgenes by ELISA and intracellular signaling as ascertained by the activation of AKT and ERK1/2 were measured; cell invasiveness was determined using a Boyden chamber assay; and the effects of the treatment were measured in vivo in a mouse model. RESULTS: In the present work, we show that the combined treatment with tGAS1 and PTEN-L inhibits the growth of pancreatic cancer cells, by reducing the activities of both AKT and ERK 1/2, decreases cell invasiveness, and restrains tumor growth in a mouse model. CONCLUSION: The combined administration of tGAS1 and PTEN-L could be a valuable adjunct therapy for the treatment of pancreatic cancer.
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INTRODUCTION AND OBJECTIVES: Curcumin, a polyphenol, is a natural compound that has been widely studied as a hepatoprotector; however, only a few studies have examined its ability to reduce fibrosis in previously established cirrhosis. The objective of this study was to investigate whether curcumin could reduce carbon tetrachloride (CCl4)-induced fibrosis and if so, to determine the action mechanisms involved in the reduction process. MATERIALS AND METHODS: CCl4 was administered to male Wistar rats (400â¯mg/kg, three times a week, i. p.) for 12 weeks; curcumin (100â¯mg/kg body weight twice per day, p. o.) was administered from week 9-12 of CCl4 treatment. Biochemical markers of hepatic injury and oxidative stress were evaluated. Hematoxylin and eosin, Masson's trichrome stains, transmission electron microscopy; immunohistochemistry, and zymography assays were carried out. Moreover, Smad3 and α-SMA mRNA and protein levels were studied. Western blotting by TGF-ß, CTGF, Col-I, MMP-13, NF-κB, IL-1, IL-10, Smad7, pSmad3, and pJNK proteins was developed. RESULTS AND CONCLUSIONS: Curcumin reduced liver damage, oxidative stress, fibrosis, and restored normal activity of MMP-9 and MMP-2. Besides, curcumin restored NF-κB, IL-1, IL-10, TGF-ß, CTGF, Col-I, MMP-13, and Smad7 protein levels. On the other hand, curcumin decreased JNK and Smad3 phosphorylation. Furthermore, curcumin treatment decreased α-SMA and Smad3 protein and mRNA levels. Curcumin normalized GSH, and NF-κB, JNK-Smad3, and TGF-ß-Smad3 pathways, leading to a decrement in activated hepatic stellate cells, thereby producing its antifibrotic effects.
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Transdiferenciação Celular/efeitos dos fármacos , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Curcumina/farmacologia , Células Estreladas do Fígado/efeitos dos fármacos , Cirrose Hepática Experimental/prevenção & controle , Fígado/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Proteína Smad3/metabolismo , Proteína Smad7/metabolismo , Animais , Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Tetracloreto de Carbono , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/patologia , Citocinas/metabolismo , Células Estreladas do Fígado/metabolismo , Células Estreladas do Fígado/ultraestrutura , Fígado/metabolismo , Fígado/ultraestrutura , Cirrose Hepática Experimental/induzido quimicamente , Cirrose Hepática Experimental/metabolismo , Cirrose Hepática Experimental/patologia , Masculino , Estresse Oxidativo/efeitos dos fármacos , Fosforilação , Ratos Wistar , Transdução de SinaisRESUMO
Glioblastoma (GBM) is the most aggressive local brain tumor and effective treatments are lacking. Many studies have proposed an important participation of steroid hormones in the development of gliomas. Evidence was provided by statistics analysis where the incidence in adult population is 50% higher in men than in women. Female patients have a better prognosis for survival compared to male patients with GBM. Also, the expression of receptors to estrogen, progesterone and androgens in glioma cell lines and tumor biopsies, and glucocorticoid receptors in GBM cell lines had been reported. Here we have investigated the effect of the pharmacological inhibition of 5-α reductases on the capacity of GBM derived cell lines C6 (rat) and U87 (human) to synthesize neurosteroids. As the knowledge of the pathways used to synthesize neurosteroids by GBM derived cells was incomplete, we have investigated the synthesis of these steroids by C6 and U87 cells using tritiated precursors and thin layer chromatography (TLC). Increasing concentrations of finasteride and dutasteride were added to U87 culture media that was collected after 24 and 48 h. The results of the study showed that C6 cells incubated with 3H-cholesterol yielded dihydroandrosterone, hydroxytestosterone, androstenediol, androstenedione and estriol, while U87 cells also synthesized progesterone, and androstanedione. Incubation with 3H-androstenedione or 3H-testosterone mainly yielded dihydrotestosterone, androsterone, dihydroandrosterone, hydroxytestosterone, and estradiol in both lines. To note, we showed here for the first time that U87 cells synthesize corticosteroids. Addition of finasteride or dutasteride to U87 cells reduced androgen and estrogen synthesis. Dutasteride also decreased the synthesis of dihydrocorticosterone and allotetrahydrodesoxycorticosterone while deoxycorticosterone was accumulated. In summary, both GBM cell lines synthesize numerous neurosteroids, including 5-α reductase products and 3α-HSD pathways that were inhibited by finasteride and dutasteride. These inhibitors may be considered as tools to control neurosteroid synthesis of potential relevance for GBM survival.
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Dutasterida/farmacologia , Finasterida/farmacologia , Glioblastoma/tratamento farmacológico , Neuroesteroides/antagonistas & inibidores , Animais , Relação Dose-Resposta a Droga , Glioblastoma/metabolismo , Glioblastoma/patologia , Humanos , Neuroesteroides/química , Neuroesteroides/metabolismo , Ratos , Células Tumorais CultivadasRESUMO
There is no effective treatment for hepatic fibrosis. Previously, we demonstrated that naringenin possesses the ability to prevent experimental chronic liver damage. Therefore, the objective of this work was to investigate whether naringenin could reverse carbon tetrachloride (CCl4)-induced fibrosis in rats and, if so, to search for the mechanisms involved. CCl4 was given to male Wistar rats (400â¯mg/kg, three times per week, i. p.) for 12 weeks; naringenin (100â¯mg/kg twice per day, p. o.) was administered from weeks 9-12 of the CCl4 treatment. Liver damage and oxidative stress markers were measured. Masson's trichrome, hematoxylin-eosin staining and immunohistochemistry were performed. Zymography assays for MMP-9 and MMP-2 were carried out. TGF-ß, CTGF, Col-I, MMP-13, NF-κB, IL-1ß, IL-10, Smad7, pSmad3 and pJNK protein levels were determined by western blotting. In addition, α-SMA and Smad3 protein and mRNA levels were studied. Naringenin reversed liver damage, biochemical and oxidative stress marker elevation, and fibrosis and restored normal MMP-9 and MMP-2 activity. The flavonoid also preserved NF-κB, IL-1ß, IL-10, TGF-ß, CTGF, Col-I, MMP-13 and Smad7 protein levels. Moreover, naringenin decreased JNK activation and Smad3 phosphorylation in the linker region. Finally, α-SMA and Smad3 protein and mRNA levels were reduced by naringenin administration. The results of this study demonstrate that naringenin blocks oxidative stress, inflammation and the TGF-ß-Smad3 and JNK-Smad3 pathways, thereby carrying out its antifibrotic effects and making it a good candidate to treat human fibrosis, as previously demonstrated in toxicological and clinical studies.
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Progressão da Doença , Flavanonas/farmacologia , Células Estreladas do Fígado/efeitos dos fármacos , Células Estreladas do Fígado/patologia , Cirrose Hepática/patologia , Animais , Biomarcadores/metabolismo , Diferenciação Celular/efeitos dos fármacos , Colágeno/metabolismo , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/metabolismo , Flavanonas/uso terapêutico , Cirrose Hepática/tratamento farmacológico , Masculino , Proteólise/efeitos dos fármacos , Ratos , Ratos Wistar , Transdução de Sinais/efeitos dos fármacosRESUMO
Parkin (PRKN) is a ubiquitin E3 ligase that catalyzes the ubiquitination of several proteins. Mutations in the human Parkin gene, PRKN, leads to degeneration of dopaminergic (DA) neurons, resulting in autosomal recessive early-onset parkinsonism and the loss of PRKN function is linked to sporadic Parkinson's disease (PD). Additionally, several in vitro studies have shown that overexpression of exogenous PRKN protects against the neurotoxic effects induced by a wide range of cellular stressors, emphasizing the need to study the mechanism(s) governing PRKN expression and induction. Here, Prkn was identified as a novel target gene of the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor and member of the bHLH/PAS (basic helix-loop-helix/Per-Arnt-Sim) superfamily. AhR binds and transactivates the Prkn gene promoter. We also demonstrated that AhR is expressed in DA neurons and that its activation upregulates Prkn mRNA and protein levels in the mouse ventral midbrain. Additionally, the AhR-dependent increase in PRKN levels is associated with a decrease in the protein levels of its target substrate, α-synuclein, in an AhR-dependent manner, because this effect is not observed in Ahr-null mice. These results suggest that treatments designed to induce PRKN expression through the use of nontoxic AhR agonist ligands may be novel strategies to prevent and delay PD.
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Ubiquitina-Proteína Ligases/metabolismo , alfa-Sinucleína/metabolismo , Actinas/metabolismo , Animais , Encéfalo/metabolismo , Linhagem Celular , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Regulação da Expressão Gênica/fisiologia , Humanos , Fígado/metabolismo , Camundongos , Camundongos Knockout , Neurônios/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Hidrocarboneto Arílico/metabolismo , Ubiquitina-Proteína Ligases/genética , alfa-Sinucleína/genéticaRESUMO
Antecedentes: la primera colecistectomía fue realizada por Carl Langenbuch en julio de 1882. Erich Mühe realizó la primera colecistectomía laparoscópica en 1982. Objetivos: evaluar la morbimortalidad de la colecistectomía realizada por residentes en nuestro Servicio. Material y métodos: trabajo retrospectivo transversal. Se tomaron los datos de las historias clínicas y los libros de quirófano de pacientes intervenidos quirúrgicamente en el hospital José Ramón Vidal de Corrientes entre enero de 2012 y enero de 2015. Se realizaron 1870 colecistectomías: 1292 (69,09%) por abordaje laparoscópico y 578 (30,58%) por abordaje convencional. Todos los procedimientos fueron iniciados y finalizados por médicos residentes bajo supervisión de un médico del staff o jefe de residentes. Resultados: 1203 (64,33%) fueron pacientes femeninos y 667 (35,66%), masculinos. En 10 pacientes hubo necesidad de conversión, siendo la tasa del 0,77%. La morbilidad fue del 0,53% (10 pacientes), la mortalidad fue del 0,05% (1 paciente). Los residentes de segundo año realizaron 240 (12,83%) procedimientos, los de tercero 1016 (54,33%) y los de cuarto año realizaron 311(16,63%) colecistectomías. En el 96,84% (1811) de los pacientes la internación en el posoperatorio fue de 24 horas. En el resto, 59 (3,15%) pacientes, la internación fue de 2 días. Conclusión: creemos que la colecistectomía convencional y la laparoscópica son de gran importancia en la formación del residente de cirugía; se trata de un procedimiento que puede y tiene que ser realizado por ellos, con supervisión apropiada, en un programa adecuado de sistema de residencias médicas.
Background: The first cholecystectomy was performed by Carl Langenbuch in July 1882. Erich Mühe performed the first laparoscopic cholecystectomy in 1982. Objectives: The aim of this study was to evaluate morbidity and mortality associated with cholecystectomy performed by residents in our service. Methods: We conducted a retrospective cross-sectional study. The information was retrieved from the medical records and operating room records of patients undergoing surgery at the Hospital José Ramón Vidal between January 2012 and January 2015. A total of 1870 cholecystectomies were performed: 1292 (69.1%) were laparoscopic procedures and 578 (30.658%) by conventional approach. All procedures were started and completed by resident physicians under the strict supervision of a staff physician or chief resident. Results: 1203 (64.33%) patients were women and 667 (35.66%) were men. Ten patients (0.77%) required conversion. Morbidity was 0.53% (10 patients) and mortality was 0.05% (one patient). Second-year residents performed 240 (12.83%) procedures, third-year residents performed 1016 (54,33%) and fourth-year residents 311 (16.63%). After surgery, 1881 (96.8%) patients remained hospitalized for 24 hours and hospital stay duration was of two days in 59 (3.15%) patients. Conclusions: Conventional and laparoscopic cholecystectomy are of great importance for the comprehensive training of residents in surgery: therefore, they constitute a procedure that can and must be performed by them, with appropriate supervision of a staff physician in adequate residency programs.
Assuntos
Humanos , Masculino , Feminino , Colecistectomia/mortalidade , Indicadores de Morbimortalidade , Argentina , Estudos Transversais , Estudos Retrospectivos , Vesícula Biliar , Corpo Clínico HospitalarRESUMO
The overexpression of GAS1 (Growth Arrest Specific 1) in glioma cells induces cell cycle arrest and apoptosis. We previously demonstrated that the apoptotic process set off by GAS1 is caused by its capacity to inhibit the Glial cell-derived neurotrophic factor (GDNF)-mediated intracellular survival signaling pathway. Whereas on the other hand, PTEN is a tumor suppressor, inactive in many tumors, and both GAS1 and PTEN inhibit the PI3K/AKT pathway. Therefore, it is relevant to investigate the potential additive effect of the overexpression of GAS1 and PTEN on tumor growth. In particular, we employed secreted forms of both GAS1 (tGAS1) and PTEN (PTEN-LONG, or PTEN-L) and tested their combined effect on glioma cells. We observed that the co-expression of both the proteins inhibited the growth of U-87 MG human glioblastoma cells more effectively than when independently expressed, and decreased the activity of both AKT and ERK1/2. Interestingly, the combination of the soluble forms was always the most effective treatment. To improve the transfer of tGAS1 and PTEN-L, we employed a lentiviral vector with a p2A peptide-enabled dual expression system that allowed the generation of the two proteins using a single promoter (CMV), in equimolar amounts. The viral vector reduced the growth of U-87 MG cells in vitro and had a striking effect in inhibiting their proliferation after inoculating it into the immunosuppressed mice. The present results support a potential adjuvant role for the combined use of tGAS1 and PTEN-L in the treatment of glioblastoma.
Assuntos
Proteínas de Ciclo Celular/genética , Vetores Genéticos/administração & dosagem , Glioblastoma/genética , PTEN Fosfo-Hidrolase/genética , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Proteínas de Ciclo Celular/administração & dosagem , Proliferação de Células/efeitos dos fármacos , Proteínas Ligadas por GPI/administração & dosagem , Proteínas Ligadas por GPI/genética , Regulação Neoplásica da Expressão Gênica , Vetores Genéticos/imunologia , Fator Neurotrófico Derivado de Linhagem de Célula Glial/genética , Glioblastoma/imunologia , Glioblastoma/patologia , Glioblastoma/terapia , Humanos , Lentivirus/genética , Camundongos , PTEN Fosfo-Hidrolase/administração & dosagem , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Growth arrest-specific 1 (Gas1) is a pleiotropic protein that induces apoptosis of tumor cells and has important roles during development. Recently, the presence of two forms of Gas1 was reported: one attached to the cell membrane by a GPI anchor; and a soluble extracellular form shed by cells. Previously, we showed that Gas1 is expressed in different areas of the adult mouse CNS. Here, we report the levels of Gas1 mRNA protein in different regions and analyzed its expressions in glutamatergic, GABAergic, and dopaminergic neurons. We found that Gas1 is expressed in GABAergic and glutamatergic neurons in the Purkinje-molecular layer of the cerebellum, hippocampus, thalamus, and fastigial nucleus, as well as in dopaminergic neurons of the substantia nigra. In all cases, Gas1 was found in the cell bodies, but not in the neuropil. The Purkinje and the molecular layers show the highest levels of Gas1, whereas the granule cell layer has low levels. Moreover, we detected the expression and release of Gas1 from primary cultures of Purkinje cells and from hippocampal neurons as well as from neuronal cell lines, but not from cerebellar granular cells. In addition, using SH-SY5Y cells differentiated with retinoic acid as a neuronal model, we found that extracellular Gas1 promotes neurite outgrowth, increases the levels of tyrosine hydroxylase, and stimulates the inhibition of GSK3ß. These findings demonstrate that Gas1 is expressed and released by neurons and promotes differentiation, suggesting an important role for Gas1 in cellular signaling in the CNS.
Assuntos
Encéfalo/metabolismo , Proteínas de Ciclo Celular/metabolismo , Diferenciação Celular/fisiologia , Neurônios/metabolismo , Animais , Neurônios Dopaminérgicos/metabolismo , Proteínas Ligadas por GPI/metabolismo , Ácido Glutâmico/metabolismo , Masculino , Camundongos , Substância Negra/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismo , Ácido gama-Aminobutírico/metabolismoRESUMO
Adverse gastrointestinal (GI) effects caused by nonsteroidal anti-inflammatory drugs (NSAIDs), including indomethacin, are recognized as the major limitation to their clinical use. NSAID-induced gastric damage is generated by cyclooxygenase inhibition, activation of inflammatory processes, and oxidative stress. Docosahexaenoic acid (DHA), an omega-3 polyunsaturated fatty acid, has shown gastroprotective effects; however, the molecular mechanisms underlying these effects have not been fully explained. As a result, the aim of this study was to examine DHA's anti-inflammatory and antioxidative actions in a mouse model of indomethacin-induced gastric injury. Oral administration of DHA (3, 10, 30, and 100mg/kg) caused a reduction in indomethacin-induced gastric hemorrhagic lesions. We found that the gastroprotective effects of DHA treatment (100mg/kg) were accompanied by decreases in several parameters: in leukocyte recruitment; gastric levels of myeloperoxidase; leukotriene B4; intercellular adhesion molecule-1; tumor necrosis factor alpha; and nuclear translocation of nuclear factor-кB. Concurrently, we observed an improvement in antioxidant defenses produced by the increase in superoxide dismutase and glutathione activities but not catalase; in addition, a decrease in some oxidative damage markers such as malondialdehyde and carbonyl proteins in lipids and proteins was observed. Furthermore, resolvin D1 production and expression of free fatty acid receptor 4 were stimulated by DHA. Therefore, this study identified the antioxidant and anti-inflammatory actions of DHA as the main mechanisms involved in DHA's gastroprotective effects against indomethacin-induced gastric damage.
Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Ácidos Docosa-Hexaenoicos/farmacologia , Indometacina/efeitos adversos , Estômago/efeitos dos fármacos , Estômago/lesões , Animais , Citoproteção/efeitos dos fármacos , Mucosa Gástrica/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Receptores Acoplados a Proteínas G/metabolismo , Estômago/citologiaRESUMO
AIM: To study the molecular mechanisms involved in the hepatoprotective effects of naringenin (NAR) on carbon tetrachloride (CCl4)-induced liver fibrosis. METHODS: Thirty-two male Wistar rats (120-150 g) were randomly divided into four groups: (1) a control group (n = 8) that received 0.7% carboxy methyl-cellulose (NAR vehicle) 1 mL/daily p.o.; (2) a CCl4 group (n = 8) that received 400 mg of CCl4/kg body weight i.p. 3 times a week for 8 wk; (3) a CCl4 + NAR (n = 8) group that received 400 mg of CCl4/kg body weight i.p. 3 times a week for 8 wk and 100 mg of NAR/kg body weight daily for 8 wk p.o.; and (4) an NAR group (n = 8) that received 100 mg of NAR/kg body weight daily for 8 wk p.o. After the experimental period, animals were sacrificed under ketamine and xylazine anesthesia. Liver damage markers such as alanine aminotransferase (ALT), alkaline phosphatase (AP), γ-glutamyl transpeptidase (γ-GTP), reduced glutathione (GSH), glycogen content, lipid peroxidation (LPO) and collagen content were measured. The enzymatic activity of glutathione peroxidase (GPx) was assessed. Liver histopathology was performed utilizing Masson's trichrome and hematoxylin-eosin stains. Zymography assays for MMP-9 and MMP-2 were carried out. Hepatic TGF-ß, α-SMA, CTGF, Col-I, MMP-13, NF-κB, IL-1, IL-10, Smad7, Smad3, pSmad3 and pJNK proteins were detected via western blot. RESULTS: NAR administration prevented increases in ALT, AP, γ-GTP, and GPx enzymatic activity; depletion of GSH and glycogen; and increases in LPO and collagen produced by chronic CCl4 intoxication (P < 0.05). Liver histopathology showed a decrease in collagen deposition when rats received NAR in addition to CCl4. Although zymography assays showed that CCl4 produced an increase in MMP-9 and MMP-2 gelatinase activity; interestingly, NAR administration was associated with normal MMP-9 and MMP-2 activity (P < 0.05). The anti-inflammatory, antinecrotic and antifibrotic effects of NAR may be attributed to its ability to prevent NF-κB activation and the subsequent production of IL-1 and IL-10 (P < 0.05). NAR completely prevented the increase in TGF-ß, α-SMA, CTGF, Col-1, and MMP-13 proteins compared with the CCl4-treated group (P < 0.05). NAR prevented Smad3 phosphorylation in the linker region by JNK since this flavonoid blocked this kinase (P < 0.05). CONCLUSION: NAR prevents CCl4 induced liver inflammation, necrosis and fibrosis, due to its antioxidant capacity as a free radical inhibitor and by inhibiting the NF-κB, TGF-ß-Smad3 and JNK-Smad3 pathways.