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2.
ACS Appl Polym Mater ; 5(11): 9612-9619, 2023 Nov 10.
Article in English | MEDLINE | ID: mdl-37970530

ABSTRACT

In the present work, we demonstrate the formation of oxide porous and nanodot structures from the same block copolymer (BCP) by the phase inversion of a BCP template. We investigated the effect of solvent annealing time on the ordering of asymmetric, cylinder forming, polystyrene-b-poly(4-vinylpyridine) (PS-b-P4VP) BCP. Phase separation of PS-b-P4VP was achieved by solvent vapor annealing (SVA) in a solvent atmosphere that is (partially) selective to P4VP to initially generate hexagonally arranged, cylindrical arrays of the expected structure. The morphology of the BCP changed from P4VP hexagonally packed cylinders to an 'inverse' structure with PS cylinders embedded in a P4VP matrix. This suggests that selective swelling occurs over time such that the swollen P4VP phase becomes the majority volume component. Metal ions (Ga3+, In3+) were infiltrated into the BCP templates by a solution-mediated infiltration approach, followed by an ultraviolet-ozone treatment to remove the polymer and oxidize the metallic ions to their oxides. The findings show that a single BCP can be used to create both metal oxide arrays and porous structures of metal oxides by simply varying the duration of the solvent annealing process. The resulting structures were analyzed through several methods including scanning electron microscopy, atomic force microscopy, X-ray photoelectron spectroscopy (XPS), transmission electron microscopy, and energy-dispersive X-ray spectroscopy. XPS analyses confirmed the complete elimination of the BCP template and the presence of metal oxides. This study provides important insights into the development of functional BCP materials with inverse structures.

3.
Polymers (Basel) ; 15(17)2023 Sep 04.
Article in English | MEDLINE | ID: mdl-37688272

ABSTRACT

The development of packaging films made from renewable raw materials, which cause low environmental impact, has gained attention due to their attractive properties, which have become an exciting option for synthetic films. In this study, cellulose micro/nanofibrils (MFC/NFC) films were produced with forest residues from the Amazon region and evaluated for their potential to generate alternative packaging to traditional plastic packaging. The MFC/NFC were obtained by mechanical fibrillation from fibers of açaí seeds (Euterpe oleracea), titica vine (Heteropsis flexuosa), and commercial pulps of Eucalyptus sp. for comparison. The fibrillation of the titica vine culminated in higher energy expenditure on raw materials. The açaí films showed a higher tensile strength (97.2 MPa) compared to the titica films (46.2 MPa), which also showed a higher permeability rate (637.3 g day-1 m-2). Films of all raw materials scored the highest in the grease resistance test (n° 12). The films produced in the study showed potential for use in packaging for light and low moisture products due to their adequate physical, mechanical, and barrier characteristics. New types of pre-treatments or fibrillation methods ecologically correct and viable for reducing energy consumption must be developed, mainly for a greater success of titica vine fibrillation at the nanoscale.

4.
Article in English | MEDLINE | ID: mdl-36078783

ABSTRACT

During clinical rotations, medical students experience situations in which the patients' right to privacy may be violated. The aim of this study is to analyze medical students' perception of clinical situations that affect patients' right to privacy, and to look for the influential factors that may contribute to the infringement on their rights, such as the students' age, sex, academic year or parents' educational level. A cross-sectional study was conducted with a survey via "Google Drive". It consisted of 16 questions about personal information, 24 questions about their experience when rotating and 21 questions about their opinion concerning several situations related to the right to privacy. A total of 129 medical students from various Spanish medical schools participated. Only 31% of 3rd-6th year students declared having signed a confidentiality agreement when starting their clinical practice, and most students (52%) reported that doctors "sometimes", "rarely" or "never" introduce themselves and the students when entering the patients' rooms. Additionally, about 50% of all students reported that they would take a picture of a patient's hospitalization report without his/her (consent), which would be useful for an assignment. Important mistakes during medical students' rotations have been observed, as well as a general lack of knowledge regarding patient's right to privacy among Spanish medical students. Men and older students showed better knowledge of current legislation, as well as those whose parents were both university-educated and those in higher academic years.


Subject(s)
Privacy , Students, Medical , Confidentiality , Cross-Sectional Studies , Female , Humans , Male , Patient Rights , Perception
5.
Preprint in English | medRxiv | ID: ppmedrxiv-22272946

ABSTRACT

BackgroundNon-pharmaceutical interventions (NPI) play a key role in managing epidemics, yet it is challenging to evaluate their impacts on disease spread and outcomes. MethodsTo estimate the effect of a mask-wearing intervention to mitigate the spread of SARS-CoV-2 on the island of Ireland, we focused on the potential for interindividual infectious contact over time as the outcome. This is difficult to measure directly; in a companion paper we estimated it using a multi-strain epidemiological model. We used data on mask-wearing and mobility in both Northern Ireland (NI) and the Republic of Ireland (ROI) to predict independently the estimated infectious contact over time. We made counterfactual predictions of infectious contact rates and hospitalisations under a hypothetical intervention where 90% of the population were wearing masks during early 2020, when in reality few people were wearing masks in public; this was mandated in both jurisdictions on 10th August 2020. ResultsThere were 1601 hospitalisations with COVID-19 in NI between 12th March and 10th August 2020, and 1521 in ROI between 3rd April and 10th August 2020. Under the counterfactual mask-wearing scenario, we estimated 512 (95% CI 400, 730) hospitalisations in NI, and 344 (95% CI 266, 526) in ROI, during the same periods. ConclusionsWe have estimated a large effect of population mask-wearing on COVID-19 hospitalisations. This could be partly due to other factors that were also changing over time.

6.
Preprint in English | medRxiv | ID: ppmedrxiv-22272942

ABSTRACT

Mathematical modelling plays a key role in understanding and predicting the epidemiological dynamics of infectious diseases. We construct a flexible discrete-time model that incorporates multiple viral strains with different transmissibilities to estimate the changing infectious contact that generates new infections. Using a Bayesian approach, we fit the model to longitudinal data on hospitalisation with COVID-19 from the Republic of Ireland and Northern Ireland during the first year of the pandemic. We describe the estimated change in infectious contact in the context of governmentmandated non-pharmaceutical interventions in the two jurisdictions on the island of Ireland. We take advantage of the fitted model to conduct counterfactual analyses exploring the impact of lockdown timing and introducing a novel, more transmissible variant. We found substantial differences in infectious contact between the two jurisdictions during periods of varied restriction easing and December holidays. Our counterfactual analyses reveal that implementing lockdowns earlier would have decreased subsequent hospitalisation substantially in most, but not all cases, and that an introduction of a more transmissible variant - without necessarily being more severe - can cause a large impact on the health care burden.

7.
United European Gastroenterol J ; 9(6): 718-726, 2021 07.
Article in English | MEDLINE | ID: mdl-34077636

ABSTRACT

INTRODUCTION: Endoscopic submucosal dissection (ESD) has become the treatment of choice for early gastric malignancies. In recent years, the ESD technique has been implemented in Western countries with increasing use. OBJECTIVES: To describe the results of gastric ESD in a Western country with a low incidence of gastric cancer. PATIENTS AND METHODS: The prospective national registry was conducted over 4 years in 23 hospitals, including 30 endoscopists. Epithelial and subepithelial lesions (SEL) qualified to complete removal with ESD were assessed. The technique, instruments, and solution for submucosal injection varied at the endoscopist's discretion. ESD was defined as difficult when: en-bloc resection was not achieved, had to be converted to a hybrid resection, lasted more than 2 h or an intraprocedural perforation occurred. Additionally, independent risk factors for difficult ESD were analyzed. RESULTS: Two hundred and thirty gastric ESD in 225 patients were performed from January 2016 to December 2019 (196 epithelial and 34 SEL). Most lesions were located in the lower stomach (111; 48.3%). One hundred and twenty-eight (55.6%) ESD were considered difficult. The median procedure time was 105 min (interquartile range [IQR]: 60-150). The procedure time for SEL was shorter than for epithelial lesions (90 min [45-121] vs. 110 min [62-160]; p = 0.038). En-bloc, R0, and curative resection rates were 91.3%, 75.2%, and 70.9%, respectively. Difficult ESD had lower R0 resection rates than ESD that did not meet the difficulty criteria (64.8% and 87.6%; p = 0.000, respectively). Fibrosis and poor maneuverability were independent factors associated with difficult ESD (OR 3.6, 95%CI 1.1-11.74 and OR 5.07, 95%CI 1.6-16.08; respectively). CONCLUSIONS: Although the number of cases is limited, the results of this analysis show acceptable en-bloc and R0 rates in gastric ESD considering the wide variability in experience among the operators. Fibrosis and poor maneuverability were associated with more difficulty in completing ESD.


Subject(s)
Endoscopic Mucosal Resection/methods , Gastric Mucosa/pathology , Gastric Mucosa/surgery , Stomach Neoplasms/pathology , Stomach Neoplasms/surgery , Adult , Aged , Aged, 80 and over , Endoscopy, Gastrointestinal , Female , Fibrosis , Humans , Incidence , Logistic Models , Male , Middle Aged , Prospective Studies , Registries , Spain/epidemiology , Stomach Neoplasms/epidemiology , Treatment Outcome
8.
Preprint in English | medRxiv | ID: ppmedrxiv-20165480

ABSTRACT

IntroductionCOVID-19 affected worldwide, causing to date, around 500,000 deaths. In Mexico, by April 29, the general case fatality was 6.52%, with 11.1% confirmed case mortality and hospital recovery rate around 72%. Once hospitalized, the odds for recovery and hospital death rates depend mainly on the patients comorbidities and age. In Mexico, triage guidelines use algorithms and risk estimation tools for severity assessment and decision-making. The studys objective is to analyze the underlying conditions of patients hospitalized for COVID-19 in Mexico concerning four severity outcomes. Materials and MethodsRetrospective cohort based on registries of all laboratory-confirmed patients with the COVID-19 infection that required hospitalization in Mexico. Independent variables were comorbidities and clinical manifestations. Dependent variables were four possible severity outcomes(a) pneumonia, (b) mechanical ventilation (c) intensive care unit, and (d) death; all of them were coded as binary Results: We included 69,334 hospitalizations of laboratory-confirmed and hospitalized patients to June 30, 2020. Patients were 55.29 years, and 62.61% were male. Hospital mortality among patients aged<15 was 9.11%, 51.99% of those aged >65 died. Male gender and increasing age predicted every severity outcome. Diabetes and hypertension predicted every severity outcome significantly. Obesity did not predict mortality, but CKD, respiratory diseases, cardiopathies were significant predictors. ConclusionObesity increased the risk for pneumonia, mechanical ventilation, and intensive care admittance, but it was not a predictor of in-hospital death. Patients with respiratory diseases were less prone to develop pneumonia, to receive mechanical ventilation and intensive care unit assistance, but they were at higher risk of in-hospital death.

9.
Sci Rep ; 10(1): 5322, 2020 03 24.
Article in English | MEDLINE | ID: mdl-32210246

ABSTRACT

Sleep apnea is a sleep disorder that occurs when the breathing of a person is interrupted during the sleep. This interruption occurs because of the patient has narrowed airways and the upper airways muscles relax, closes in and blocks the airway. Therefore, any forces or reaction originated by the air flow dynamics over the relaxed upper airways muscles could make to close the upper airways, and consequently the air could not flow into your lungs, provoking sleep apnea. Fully describing the dynamic behavior of the airflow in this area is a severe challenge for the physicians. In this paper we explore the dynamic behavior of airflow in the upper airways of 6 patients suffering obstructive sleep apnea with/without a mandibular advancement device using computational fluid dynamics. The development of flow unsteadiness from a laminar state at entry to the pharynx through to the turbulent character in the soft palate area is resolved using an accurate numerical model. Combining the airflow solution with a geometrical analysis of the upper airways reveals the positive effects of mandibular advance device in the air flow behavior (pressure drop). Improved modeling of airflow and positioning of mandibular advance device could be applied to improve diagnosis and treatment of obstructive sleep apnea.


Subject(s)
Pulmonary Ventilation/physiology , Sleep Apnea Syndromes/physiopathology , Sleep Apnea Syndromes/therapy , Adult , Computer Simulation , Female , Humans , Hydrodynamics , Larynx/physiopathology , Male , Mandible/physiopathology , Middle Aged , Nose/physiopathology , Palate, Soft/physiopathology , Pharynx/physiopathology , Polysomnography , Respiration , Sleep Apnea, Obstructive/physiopathology
10.
Movimento (Porto Alegre) ; 26: e26025, 2020.
Article in Spanish | LILACS | ID: biblio-1356510

ABSTRACT

Resumen: Esta investigación tiene como objetivos, por un lado, explorar las percepciones de las deportistas acerca de cómo son evaluadas por su entorno social con relación a la feminidad y a su orientación sexual por su vinculación con una práctica deportiva de alto rendimiento. Por otro, analizar la visibilidad y aceptación de la homosexualidad en el contexto deportivo tanto por las deportistas como por los agentes sociales que las rodean. La metodología fue de tipo cualitativo. Se llevaron a cabo cinco grupos focales en los que participaron 46 mujeres practicantes de futbol sala, balonmano, voleibol, atletismo y natación. Los resultados muestran que las deportistas presentan un discurso de resistencia frente al estereotipo de marimacho-lesbiana que, comprobamos, continúa vigente en el imaginario colectivo. A pesar de que se perciben avances sociales en este sentido, la homosexualidad sigue estigmatizada y aún persisten formas de rechazo cognitivo-afectivo en parte de la sociedad española.


Resumo: Esta pesquisa tem como objetivo, por um lado, explorar as percepções das atletas sobre como são avaliadas pelo seu ambiente social em relação à feminilidade e sua orientação sexual, para vincular-se a uma prática esportiva de alto desempenho e, por outro, analisar a visibilidade e aceitação da homossexualidade no contexto esportivo, tanto pelos atletas como pelos agentes sociais ao seu redor. A metodologia foi qualitativa. Foram feitos cinco grupos focais nos quais participaram 46 mulheres praticando futsal, handebol, vôlei, atletismo e natação. Os resultados mostram que as atletas apresentam um discurso de resistência contra o estereótipo da "Maria-João" o "sapatão" que, verificamos, ainda está em vigor na imaginação coletiva. Embora os avanços sociais sejam percebidos nesse sentido, a homossexualidade permanece estigmatizada e ainda persistem as formas de rejeição cognitivo-afetiva em parte da sociedade espanhola.


Abstract: On the one hand, this research aims to explore the perceptions of women practicing high-performance sports about how they are viewed by their social environment in terms of their femininity and sexual orientation. On the other hand, it analyzes the visibility and acceptance of homosexuality in the sports context by both athletes and the social agents surrounding them. The research methodology was qualitative, with five focus groups including 46 women who practiced futsal, handball, volleyball, athletics and swimming. Results show that they present a resistance-oriented discourse against butch-lesbian stereotypes that, as we prove, are still present in collective imagination. Despite social advances perceived in this regard, homosexuality remains stigmatized and forms of cognitive-affective rejection persist in part of Spanish society.


Subject(s)
Humans , Female , Track and Field , Sexuality , Femininity , Sexual and Gender Minorities , Gender Identity , Homosexuality , Qualitative Research
11.
Rev. bioét. (Impr.) ; 27(1): 53-61, jan.-mar. 2019.
Article in Spanish | LILACS | ID: biblio-1041967

ABSTRACT

Resumen La religión es un elemento de la cultura que está presente en el proceso de salud-enfermedad-atención. Esta presencia no solamente es a nivel personal, sino a nivel colectivo a través de políticas públicas de salud. Estas políticas deben pensarse para toda la población, independientemente de si creen o no creen en una determinada religión. Sin embargo, el cristianismo es una religión proselitista, en cuya historia puede verse la imposición de su visión sobre otras visiones (la Edad Media es el mejor ejemplo). En el mundo contemporáneo, la religión continúa buscando imponer sus criterios, también en el campo de la salud. La propuesta de este trabajo es que hay cuestiones éticas que subyacen en los colectivos y que hacen que este tipo de imposiciones no sea correcto en las sociedades contemporáneas, multiculturales y moralmente pluralistas.


Abstract Religion is an element of culture that is present in the process of health-illness-care. This presence is not only at a personal level, but at a collective level through public health policies. These policies should be thought for the whole population, regardless of whether they believe or do not believe in a particular religion. However, Christianity is a proselytizing religion, in whose history can be seen imposing its vision on other visions (Middle Ages is the best example). In the contemporary world, religion continues to seek to impose its criteria, also in the field of health. The proposal of this work is that there are ethical issues that underlie the collective and that make this type of impositions is not correct in contemporary societies, multicultural and morally pluralist.


Resumo Religião é elemento cultural presente no processo saúde-doença-cuidado não apenas em nível pessoal, mas coletivo, por meio de políticas de saúde pública. Essas políticas devem ser pensadas para toda a população, independentemente da crença pessoal em determinada religião. No entanto, o cristianismo é religião proselitista, em cuja história pode-se observar a imposição de sua visão sobre outras (a Idade Média é o melhor exemplo). No mundo contemporâneo, a religião continua a procurar impor seus critérios, também no campo da saúde. Este artigo propõe que existem questões éticas subjacentes ao coletivo e que esse tipo de imposição não é correto em sociedades contemporâneas, multiculturais e moralmente pluralistas.


Subject(s)
Religion and Medicine , Bioethics , Health Sciences , Health-Disease Process
13.
Microsc Res Tech ; 75(10): 1321-8, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22622998

ABSTRACT

The enigmatic rodlet cells (RCs) are characterized by conspicuous inclusions named "rodlets". They were discovered over 100 years ago and were considered as parasites but shortly afterward interpreted as endogenous cells. The RCs have been described in different tissues of marine and freshwater teleosts, but their origin and function remain unknown. This work was designed to an ultrastructural study on RCs development and distribution in intestinal epithelium of Dicentrarchus labrax. Three different stages of RCs development from early precursor cells to mature phase were observed, as well as a migration and finally an extrusion of their contents. In this study, the immature cells were found near the basal epithelium membrane. They were mainly identified by a rough endoplasmic reticulum with dilated cisternae, by developing rodlets and a thin fibrillar coat. The maturing RCs, localized in the middle zone of the epithelium, appeared to be undergoing a reorganization of the cell organelles. The mature RCs, placed near the free surface, showed a thick subplasmalemmar fibrillar coat. Most of the organelles were aggregated at the cell apex with a basally located nucleus. A cellular polarity was more evident. One of the most conspicuous features was the occurrence of mature rodlets club-sac in shape orientated toward the cell apex. Adhesive junctions between surface epithelial cells and RCs, while discharging their contents, were seen. We have connected morphological figures and distribution to different stages of development in RCs, supporting the hypothesis of their secretory function.


Subject(s)
Bass , Intestinal Mucosa/cytology , Intestines/cytology , Animals , Cell Adhesion , Cell Movement , Organelles/ultrastructure
14.
Microsc Res Tech ; 75(5): 643-9, 2012 May.
Article in English | MEDLINE | ID: mdl-22505185

ABSTRACT

The European sea bass, a member of the Moronidae family, is a food fish, considered one of the first models for the intensive breeding in salt water. It has nowadays an important and increasing presence in the international fishing markets. Sea basses are carnivorous, feeding on little fishes and invertebrates. Considering the important role of the tongue during the intraoral transport and the swallowing of food, scarce data are present in literature about its morphology. The aim of this study was to analyze the morphology of the tongue by means of scanning electron and light microscopy. Adult sea basses were obtained from the aquarium of the Sicilian Center of Experimental Ichthyiopathology of the University of Messina. The fishes were anaesthetized with MS 222 and the heads were then quickly removed and processed for the paraffin embedding and SEM processing. Three different tongue regions could be distinguished: an apex, a body, and a root. Scanning electron and light microscopy showed the presence of numerous canine-like teeth, surrounded by taste buds and numerous fungiform and conical papillae. The teeth were curved and their tips were posteriorly oriented. The results confirm, in teleosts too, the fundamental role of the tongue in the mechanics of food ingestion. Moreover, the presence of taste buds demonstrates the interaction of food processing and taste. These data could be a potential source to identify new and better methods of nutrition in the breeding of this fish.


Subject(s)
Bass/anatomy & histology , Tongue/anatomy & histology , Animals , Microscopy
15.
Brain Res ; 1442: 9-14, 2012 Mar 09.
Article in English | MEDLINE | ID: mdl-22306032

ABSTRACT

The zebrafish pineal gland plays an important role in different physiological functions including the regulation of the circadian clock. In the fish pineal gland the pinealocytes are made up of different segments: outer segment, inner segment and basal pole. Particularly, in the outer segment the rhodopsin participates in the external environment light reception that represents the first biochemical step in the melatonin production. It is well known that the rhodopsin in the adult zebrafish is well expressed in the pineal gland but both the expression and the cellular localization of this protein during development remain still unclear. In this study using qRT-PCR, sequencing and immunohistochemistry the expression as well as the protein localization of the rhodopsin in the zebrafish from larval (10 dpf) to adult stage (90 dpf) were demonstrated. The rhodopsin mRNA expression presents a peak of expression at 10 dpf, a further reduction to 50 dpf before increasing again in the adult stage. Moreover, the cellular localization of the rhodopsin-like protein was always localized in the pinealocyte at all ages examined. Our results demonstrated the involvement of the rhodopsin in the zebrafish pineal gland physiology particularly in the light capture during the zebrafish lifespan.


Subject(s)
Pineal Gland/metabolism , Rhodopsin/metabolism , Zebrafish Proteins/metabolism , Zebrafish/growth & development , Zebrafish/metabolism , Animals , Gene Expression Regulation, Developmental , Immunohistochemistry , Larva/growth & development , Larva/metabolism , Pineal Gland/cytology , Pineal Gland/growth & development , Reverse Transcriptase Polymerase Chain Reaction , Rhodopsin/genetics , Zebrafish/genetics , Zebrafish Proteins/genetics
16.
Microsc Res Tech ; 75(3): 359-66, 2012 Mar.
Article in English | MEDLINE | ID: mdl-21858901

ABSTRACT

The zebrafish pineal gland plays a fundamental role in the regulation of the circadian rhythm through the melatonin secretion. The pinealocytes, also called photoreceptive cells, are considered the morphofunctional unit of pineal gland. In literature, the anatomical features, the cellular characteristics, and the pinealocytes morphology of zebrafish pineal gland have not been previously described in detail. Therefore, this study was undertaken to analyze the structure and ultrastructure, as well as the immunohistochemical profile of the zebrafish pineal gland with particular reference to the pinealocytes. Here, we demonstrated, using RT-PCR, immunohistochemistry and transmission electron microscopy, the expression of the mRNA for rhodopsin in the pineal gland of zebrafish, as well as its cellular localization exclusively in the pinealocytes of adult zebrafish. Moreover, the ultrastructural observations demonstrated that the pinealocytes were constituted by an outer segment with numerous lamellar membranes, an inner segment with many mitochondria, and a basal pole with the synapses. Our results taken together demonstrated a central role of zebrafish pinealocytes in the control of pineal gland functions.


Subject(s)
Photoreceptor Cells, Vertebrate/ultrastructure , Pineal Gland/cytology , Zebrafish/anatomy & histology , Animals , Immunohistochemistry , Microscopy, Electron, Transmission , Reverse Transcriptase Polymerase Chain Reaction , Rhodopsin/analysis , Rhodopsin/metabolism
17.
Cytotherapy ; 13(10): 1249-55, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21999374

ABSTRACT

BACKGROUND AIMS. Lymphedema is a common complication with breast cancer treatment that does not have a definite cure. Our objective was to determine the efficacy of autologous stem cells (ASC) in the treatment of lymphedema secondary to mastectomy and axillary lymphadenectomy in comparison with traditional decongestive treatment with compression sleeves. METHODS. A prospective study including 20 women with lymphedema secondary to breast cancer surgery with axillary lymphadenectomy was conducted. Women were assigned at random to one of two groups. One group of 10 women was injected with ASC in the affected arm, whereas the other 10 women comprised the control group and received traditional compression sleeve therapy (CST). The follow-up for both groups was 12 weeks. Pain, sensitivity and mobility were assessed before and after therapy. RESULTS. There was improvement in the volume of lymphedema in both groups, with no significant difference. In the ASC group there was an overall volume reduction during the follow-up, whereas in the CST group lymphedema recurred after the compression sleeve was removed. CONCLUSIONS. Our findings suggest that ASC injection for patients with lymphedema can be an effective treatment. It reduces arm volume and associated co-morbidities of pain and decreased sensitivity. Traditional CST was also effective for lymphedema reduction, but it was dependent on continuous use of the treatment.


Subject(s)
Breast Neoplasms/therapy , Carcinoma/therapy , Lymphedema/etiology , Postoperative Complications , Stem Cell Transplantation , Aged , Breast Neoplasms/pathology , Breast Neoplasms/physiopathology , Breast Neoplasms/surgery , Carcinoma/pathology , Carcinoma/physiopathology , Carcinoma/surgery , Compression Bandages , Female , Follow-Up Studies , Humans , Lymphedema/prevention & control , Mastectomy , Middle Aged , Pilot Projects , Prospective Studies , Transplantation, Autologous
18.
J Biol Chem ; 279(53): 55682-9, 2004 Dec 31.
Article in English | MEDLINE | ID: mdl-15485846

ABSTRACT

A number of cell types express inducible nitric-oxide synthase (NOS2) in response to exogenous insults such as bacterial lipopolysaccharide or proinflammatory cytokines. Although it has been known for some time that the N-terminal end of NOS2 suffers a post-translational modification, its exact identification has remained elusive. Using radioactive fatty acids, we show herein that NOS2 becomes thioacylated at Cys-3 with palmitic acid. Site-directed mutagenesis of this single residue results in the absence of the radiolabel incorporation. Acylation of NOS2 is completely indispensable for intracellular sorting and .NO synthesis. In fact, a C3S mutant of NOS2 is completely inactive and accumulates to intracellular membranes that almost totally co-localize with the Golgi marker beta-cop. Likewise, low concentrations of the palmitoylation blocking agents 2-Br-palmitate or 8-Br-palmitate severely affected the .NO synthesis of both NOS2 induced in muscular myotubes and transfected NOS2. However, unlike endothelial NOS, palmitoylation of inducible NOS is not involved in its targeting to caveolae. We have created 16 NOS2-GFP chimeras to inspect the effect of the neighboring residues of Cys-3 on the degree of palmitoylation. In this regard, the hydrophobic residue Pro-4 and the basic residue Lys-6 seem to be indispensable for palmitoylation. In addition, agents that block the endoplasmic reticulum to Golgi transit such as brefeldin A and monensin drastically reduced NOS2 activity leading to its accumulation in perinuclear areas. In summary, palmitoylation of NOS2 at Cys-3 is required for both its activity and proper intracellular localization.


Subject(s)
Cysteine/chemistry , Nitric Oxide Synthase/metabolism , Nitric Oxide/metabolism , Palmitic Acid/metabolism , Amino Acid Sequence , Animals , Anti-Bacterial Agents/pharmacology , Antifungal Agents/pharmacology , Biological Transport , Boron Compounds/pharmacology , Brefeldin A/pharmacology , COS Cells , Cell Line , Cells, Cultured , Cloning, Molecular , Escherichia coli/metabolism , Fluorescent Dyes/pharmacology , Golgi Apparatus/metabolism , Green Fluorescent Proteins/metabolism , Hydroxylamine/chemistry , Lysine/chemistry , Mice , Molecular Sequence Data , Monensin/pharmacology , Mutagenesis, Site-Directed , Mutation , Nitric Oxide/chemistry , Nitric Oxide Synthase Type II , Palmitic Acid/chemistry , Proline/chemistry , Protein Processing, Post-Translational , Recombinant Fusion Proteins/metabolism , Recombinant Proteins/chemistry , Serine/chemistry , Time Factors , Transfection
19.
J Cell Sci ; 117(Pt 9): 1687-97, 2004 Apr 01.
Article in English | MEDLINE | ID: mdl-15075230

ABSTRACT

Several cell types express inducible nitric oxide synthase (NOS2) in response to exogenous insults such as bacterial lipopolysaccharide (LPS) or proinflammatory cytokines. For instance, muscular cells treated with LPS and interferon gamma (IFN-gamma) respond by increasing the mRNA and protein levels of NOS2, and synthesize large amounts of nitric oxide. We show here that transcriptional induction of NOS2 in muscular cells proceeds with a concomitant decrease in the levels of caveolin-1, -2 and -3. Addition of *NO-releasing compounds to C2C12 muscle cells reveals that this downregulation of the caveolin (cav) levels is due to the presence of *NO itself in the case of caveolin-3 and to the action of the LPS/IFN-gamma in the case of cav-1 and cav-2. Likewise, muscle cells obtained from NOS2(-/-) knockout mice challenged with LPS/IFN-gamma could downregulate their levels of cav-1 but not of cav-3, unlike wild-type animals, in which both cav-1 and cav-3 levels diminished in the presence of the proinflammatory insult. Laser confocal immunofluorescence analysis proves that *NO exerts autocrine and paracrine actions, hence diminishing the cav-3 levels. When the induced NOS2 was purified using an affinity resin or immunoprecipitated from muscular tissues, it appears strongly bound not only to calmodulin but also to cav-1, and marginally to cav-2 and cav-3. When the cav levels where reduced using antisense oligonucleotides, an increase in the NOS2-derived.NO levels could be measured, demonstrating the inhibitory role of the three cav isoforms. Our results show that cells expressing NOS2 diminish their cav levels when the synthesis of *NO is required.


Subject(s)
Argininosuccinic Acid/analogs & derivatives , Caveolins/metabolism , Down-Regulation , Nitric Oxide Synthase/metabolism , Amidines/pharmacology , Animals , Argininosuccinic Acid/pharmacology , Benzylamines/pharmacology , Caveolin 1 , Caveolin 2 , Caveolin 3 , Caveolins/genetics , Cell Line , Down-Regulation/drug effects , Enzyme Induction/drug effects , Enzyme Inhibitors/pharmacology , Flavonoids/pharmacology , Interferon-gamma/pharmacology , Lipopolysaccharides/pharmacology , Mice , Mice, Knockout , Muscle Cells/drug effects , Muscle Cells/metabolism , Muscle Fibers, Skeletal/metabolism , Nitric Oxide Synthase/deficiency , Nitric Oxide Synthase/genetics , Nitric Oxide Synthase Type II , Nitrites/metabolism , Oligoribonucleotides, Antisense , Protein Binding , Protein Isoforms/genetics , Protein Isoforms/metabolism
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