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1.
Cell Mol Biol (Noisy-le-grand) ; 69(13): 24-30, 2023 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-38158693

RESUMO

We focus on this study in designing an alternative technique for obtaining mesenchymal stem cells (MSCs) from residual tissue, Hoffa fat, in arthroscopic procedures. Two males and two females were included, and underwent knee arthroscopy; a sample of infrapatellar adipose tissue was obtained with basket forceps. The primary culture was made using the explant method and the culture media: DMEM-high glucose, supplemented with 10% of inactivated human allogeneic serum. All the cellular cultures remained under culture conditions for three weeks, after that by flow cytometry the cells were characterized by MSCs antibody panel: CD105, CD73 and CD90. Subsequently, in the first pass, the MSCs were cultured in commercial human chondrogenic, osteogenic and adipogenic mediums, respectively. After primary culture, we obtained on average 95,600.00 ± 7,233.26 cells/cm2, and the duplication time of MSCs isolate from Hoffa fat pad was established in 39 hours. By flow cytometry, we found that surface markers percentage for expanded MSCs (CD105, CD73, CD90) in primary culture significantly increased and its morphology was fibroblastic-like. After differentiation culture which was made in the first pass, by immunofluorescence, we obtained positive cell markers for three lineages of differentiation, adipocytes: LPL protein, osteocytes: RUNX2, Osteopontin, chondrocytes: SOX9, Aggrecan and COL2A1. We managed to isolate a significant number of MSCs from this source using an easy method to implement and minimal nutrient supplementation, with high potential for differentiation to mature mesenchymal tissues and potential use in basic experimental, preclinical and even clinical research.


Assuntos
Tecido Adiposo , Células-Tronco Mesenquimais , Masculino , Feminino , Humanos , Células Cultivadas , Diferenciação Celular , Meios de Cultura/farmacologia , Meios de Cultura/metabolismo , Células-Tronco Mesenquimais/metabolismo , Proliferação de Células
2.
Mol Biol Rep ; 49(3): 2503-2517, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35013859

RESUMO

BACKGROUND: The present review is focused on general aspects of the synovial membrane as well as specialized aspects of its cellular constituents, particularly the composition and location of synovial membrane mesenchymal stem cells (S-MSCs). S-MSC multipotency properties are currently at the center of translational medicine for the repair of multiple joint tissues, such as articular cartilage and meniscus lesions. METHODS AND RESULTS: We reviewed the results of in vitro and in vivo research on the current clinical applications of S-MSCs, surface markers, cell culture techniques, regenerative properties, and immunomodulatory mechanisms of S-MSCs as well as the practical limitations of the last twenty-five years (1996 to 2021). CONCLUSIONS: Despite the poor interest in the development of new clinical trials for the application of S-MSCs in joint tissue repair, we found evidence to support the clinical use of S-MSCs for cartilage repair. S-MSCs can be considered a valuable therapy for the treatment of repairing joint lesions.


Assuntos
Cartilagem Articular , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Diferenciação Celular , Transplante de Células-Tronco Mesenquimais/métodos , Membrana Sinovial
3.
PeerJ ; 8: e10069, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33083132

RESUMO

BACKGROUND: Next-generation sequencing (NGS) is a high-throughput technology that has become widely integrated in molecular diagnostics laboratories. Among the large diversity of NGS-based panels, the Trusight Tumor 26 (TsT26) enables the detection of low-frequency variants across 26 genes using the MiSeq platform. METHODS: We describe the inter-laboratory validation and subsequent clinical application of the panel in 399 patients presenting a range of tumor types, including gastrointestinal (GI, 29%), hematologic (18%), lung (13%), gynecological and breast (8% each), among others. RESULTS: The panel is highly accurate with a test sensitivity of 92%, and demonstrated high specificity and positive predictive values (95% and 96%, respectively). Sequencing testing was successful in two-thirds of patients, while the remaining third failed due to unsuccessful quality-control filtering. Most detected variants were observed in the TP53 (28%), KRAS (16%), APC (10%) and PIK3CA (8%) genes. Overall, 372 variants were identified, primarily distributed as missense (81%), stop gain (9%) and frameshift (7%) altered sequences and mostly reported as pathogenic (78%) and variants of uncertain significance (19%). Only 14% of patients received targeted treatment based on the variant determined by the panel. The variants most frequently observed in GI and lung tumors were: KRAS c.35G > A (p.G12D), c.35G > T (p.G12V) and c.34G > T (p.G12C). CONCLUSIONS: Prior panel validation allowed its use in the laboratory daily practice by providing several relevant and potentially targetable variants across multiple tumors. However, this study is limited by high sample inadequacy rate, raising doubts as to continuity in the clinical setting.

4.
Rev. Fac. Med. UNAM ; 61(6): 43-51, nov.-dic. 2018. tab, graf
Artigo em Espanhol | LILACS | ID: biblio-990393

RESUMO

RESUMEN En el mundo actual, las llamadas "tecnologías de fabricación por adición" o impresión 3D también llamado prototipado rápido, han trascendido las fronteras de casi todos los campos de la ciencia, y su incursión en la medicina es cada vez mayor. Es justamente en el campo médico que esta tecnología de impresión por adición ha evolucionado a la bioimpresión, que incluye un proceso de cultivo celular en laboratorio haciendo posible la formación de órganos y/o tejidos personalizados. Para la impresión tridimensional de órganos en humanos se toman muestras de un tejido o células madre del paciente, para ser cultivadas y expandidas en laboratorio para su posterior diferenciación a una línea celular específica. Para este proceso se utiliza un material sólido como andamio a temperatura ambiente con un punto de fusión conocido. En la creación de un modelo para la fabricación de un órgano o tejido en impresión 3D, se utilizan los estudios de imágenes médicas de los pacientes intentando preservar al máximo la anatomía de las estructuras que se desean reproducir. En este artículo se abordan las bases y el potencial uso de esta tecnología en el área médica.


ABSTRACT In today's world, so-called "addition manufacturing technologies" or 3D printing also called rapid prototyping have transcended the borders of almost every field of science and medicine is no exception. It is not surprising that its exploration for practical uses is increasing. In medicine, this technology of printing by addition has evolved to bioprinting, which occurs by a special process, from cells grown in a laboratory, which makes possible its transformation into a type of organs tailored to the patient. The three-dimensional impression of human organs requires take samples of tissues or stem cells from the patient, which are grown in the laboratory waiting to multiply or differentiate to other cell lines; then, to create said object, a solid material at room temperature and with a known melting point is applied layer by layer. Currently the use of this technology uses the medical images of patients trying to preserve the anatomy of the structures that they want to reproduce. In this article the bases and the potential use of this technology in the medical area will be addressed.

5.
J Mol Model ; 24(10): 276, 2018 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-30194488

RESUMO

Charge carriers (electrons and holes) are generated on the TiO2 using UV radiation; this excitation energy can be reduced by modifying the material electronic structure, for example, by doping or creating oxygen vacancies. Here, the electronic structure of a transition metal-doped anatase, bulk and surface, and their interaction with oxygen vacancies are studied using density functional theory. The visible light response of metal-doped TiO2 (101) is also determined. Transition metals generate intra-band gap states, which reduce the excitation energy but may also act as charge recombination sites. Dopants Fe, Co, and Ni remarkably enhance the visible light response due to the states in the middle of the gap. However, Co and Ni create heavier charge carriers. Our results show that Pd and Pt-doped TiO2 generate states near the valence and conduction band with a "clean" band gap (without states in the middle of the gap). Moreover, Pt-doped TiO2 maintains the charge mobility because it presents a small charge carriers mass. Hence, Pt-doped TiO2 represents the best alternative to activate TiO2 under visible light. The optical response of transition metal-doped TiO2 follows the order 3d > 4d > 5d. The oxygen vacancies reduce the response of metal-doped TiO2 to visible light because the unpaired electrons generated occupy the empty states of metal-doping. Graphical Abstract Density of states of TiO2 (101) surface doped with transition metals and oxygen vacancies.

6.
Tissue Cell ; 52: 116-123, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29857820

RESUMO

Today, regenerative medicine requires new sources of multipotent stem cells for their differentiation to chondrocytes using the mediums of differentiation available in the market. This study aimed to determine whether the Mesenchymal Stem Cells (MSCs) isolated from Mobilized Peripheral Blood (MPB) in sheep using the Granulocyte Colony-Stimulating Factor (G-CSF), have the ability of first acquire a fibroblast-like morphology after being forced out of the bone marrow niche by G-CSF and second, if the cells have the capacity to express collagen type-II α I in primary culture using a human commercial media of differentiation. Six Suffolk male sheep with age of 2 years were mobilized using G-CSF. One subcutaneous injection of 10 mcg per kilogram of bodyweight were administered every 24 h during three consecutive days. At day four, a sample of 20 mL of peripheral blood was harvested, afterwards, monocytes cells were separated by ficoll gradient. The mobilized MSCs were expanded in primary culture in DMEM medium supplemented with 10% adult sheep serum for three weeks and characterized by an antibody panel for surface markers: CD105, CD90, CD73, CD34, and CD45, before and after primary culture. Subsequently, an aliquot of cells in the first pass were cultured in a commercial human chondrogenic medium for three weeks. As a result, the percentage of surface markers for MSCs (CD105, CD90, CD73) in expanded cells in primary culture significantly increased, at the same time a decrease in the markers for hematopoietic cells (CD34 and CD45) was observed and the cells morphology was fibroblast-like. After three weeks of differentiation culture, the immunofluorescence analysis evidenced the expression of collagen-type-II. It was concluded that Mesenchymal Stem Cells isolated from mobilized peripheral blood in sheep have the ability to pre-differentiate into chondral like cells and express collagen type-II when are stimulated with a human commercial chondrogenic medium in monolayer culture.


Assuntos
Cartilagem Articular/citologia , Diferenciação Celular/efeitos dos fármacos , Células-Tronco Mesenquimais/citologia , Animais , Células Cultivadas , Meios de Cultivo Condicionados/farmacologia , Humanos , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Ovinos
7.
Acta Gastroenterol Latinoam ; 44(1): 48-51, 2014 Mar.
Artigo em Espanhol | MEDLINE | ID: mdl-24847630

RESUMO

Pneumatosis cystoides intestinalis features the presence of subserum or submucous cysts with gas in the intestinal wall. Its prevalence is 0.03%, although it has increased in the past decades due to a greater use of colonoscopy in everyday medical practice. Though there are several theories about its origin and many associated diseases were reported, its pathogenesis still remains uncertain. It is generally diagnosed as a finding in an imaging test. The treatment depends on the severity of the associated disease and symptoms. We report the case of a 59-year-old woman, heavy smoker, with no other clinical conditions who took a medical consultation due to abdominal bloating. She underwent a screening colonoscopy which detected the existence of cysts on the colonic wall. Afterwards, a computed tomography was performed and showed apical lung bullae, gas in a colonic wall segment, and ascitis associated to a big anexial tumor. She underwent a cytoreductive surgery, confirming the presence of advanced ovary neoplasm. The endoscopic biopsy confirmed the diagnosis of penumatosis cystoides intestinalis. We report these case because it is a rare entity which requires uncommon management and treatment guidelines.


Assuntos
Neoplasias Ovarianas/diagnóstico , Pneumatose Cistoide Intestinal/diagnóstico , Biópsia , Colonoscopia , Feminino , Humanos , Pessoa de Meia-Idade , Neoplasias Ovarianas/complicações , Pneumatose Cistoide Intestinal/complicações , Pneumatose Cistoide Intestinal/cirurgia , Tomografia Computadorizada por Raios X
8.
Acta gastroenterol. latinoam ; 44(1): 48-51, 2014 Mar.
Artigo em Espanhol | LILACS, BINACIS | ID: biblio-1157423

RESUMO

Pneumatosis cystoides intestinalis features the presence of subserum or submucous cysts with gas in the intestinal wall. Its prevalence is 0.03


, although it has increased in the past decades due to a greater use of colonoscopy in everyday medical practice. Though there are several theories about its origin and many associated diseases were reported, its pathogenesis still remains uncertain. It is generally diagnosed as a finding in an imaging test. The treatment depends on the severity of the associated disease and symptoms. We report the case of a 59-year-old woman, heavy smoker, with no other clinical conditions who took a medical consultation due to abdominal bloating. She underwent a screening colonoscopy which detected the existence of cysts on the colonic wall. Afterwards, a computed tomography was performed and showed apical lung bullae, gas in a colonic wall segment, and ascitis associated to a big anexial tumor. She underwent a cytoreductive surgery, confirming the presence of advanced ovary neoplasm. The endoscopic biopsy confirmed the diagnosis of penumatosis cystoides intestinalis. We report these case because it is a rare entity which requires uncommon management and treatment guidelines.


Assuntos
Neoplasias Ovarianas/diagnóstico , Pneumatose Cistoide Intestinal/diagnóstico , Biópsia , Colonoscopia , Feminino , Humanos , Neoplasias Ovarianas/complicações , Pessoa de Meia-Idade , Pneumatose Cistoide Intestinal/cirurgia , Pneumatose Cistoide Intestinal/complicações , Tomografia Computadorizada por Raios X
10.
J Chem Phys ; 131(12): 124102, 2009 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-19791847

RESUMO

In this paper we propose an iterative method for solving the inhomogeneous systems of linear equations associated with density fitting. The proposed method is based on a version of the conjugate gradient method that makes use of automatically built quasi-Newton preconditioners. The paper gives a detailed description of a parallel implementation of the new method. The computational performance of the new algorithms is analyzed by benchmark calculations on systems with up to about 35,000 auxiliary functions. Comparisons with the standard, direct approach show no significant differences in the computed solutions.

11.
Med Clin (Barc) ; 131 Suppl 3: 18-25, 2008 Dec.
Artigo em Espanhol | MEDLINE | ID: mdl-19572449

RESUMO

BACKGROUND AND OBJECTIVES: A safety culture is essential to minimize errors and adverse events. Its measurement is needed to design activities in order to improve it. This paper describes the methods and main results of a study on safety climate in a nation-wide representative sample of public hospitals of the Spanish NHS. MATERIAL AND METHOD: The Hospital Survey on Patient Safety Culture questionnaire was distributed to a random sample of health professionals in a representative sample of 24 hospitals, proportionally stratified by hospital size. Results are analyzed to provide a description of safety climate, its strengths and weaknesses. Differences by hospital size, type of health professional and service are analyzed using ANOVA. RESULTS: A total of 2503 responses are analyzed (response rate: 40%, (93% from professionals with direct patient contact). A total of 50% gave patient safety a score from 6 to 8 (on a 10-point scale); 95% reported < 2 events last year. Dimensions "Teamwork within hospital units" (71.8 [1.8]) and "Supervisor/Manager expectations and actions promoting safety" (61.8 [1.7]) have the highest percentage of positive answers. "Staffing", "Teamwork across hospital units", "Overall perceptions of safety" and "Hospital management support for patient safety" could be identified as weaknesses. Significant differences by hospital size, type of professional and service suggest a generally more positive attitude in small hospitals and Pharmacy services, and a more negative one in physicians. CONCLUSIONS: Strengths and weaknesses of the safety climate in the hospitals of the Spanish NHS have been identified and they are used to design appropriate strategies for improvement.


Assuntos
Atenção à Saúde/normas , Hospitais Públicos/normas , Cultura Organizacional , Pacientes , Gestão da Segurança , Humanos , Espanha
12.
Rev. neurocir ; 8(3): 85-90, ago.-oct. 2006. ilus
Artigo em Espanhol | LILACS | ID: lil-456135

RESUMO

El tronco encefálico se ubica en la fosa posterior, en íntima relación con el cerebelo e importantes estructuras vasculares. Contiene en su interior tractos de sustancia blanca que vinculan entre sí cerebro, tronco encefálico, médula espinal y cerebelo. Contiene en su interior los núcleos de origen de los diez últimos pares craneanos y núcleos que regulan funciones, como la mirada lateral, la deglución o incluso el ciclo sueño - vigilia. Puede ser afectado por diversos tipos de patologías, como tumores, malformaciones vasculares, procesos infecciosos, inflamatorios o desmielinizantes. El presente es el primer artículo de cuatro, destinados a analizar los abordajes quirúrgicos más frecuentes al tronco encefálico. En esta primera parte se describen aquí los principales aspectos de la anatomía de superficie del tronco cerebral.


Assuntos
Tronco Encefálico/anatomia & histologia , Tronco Encefálico/cirurgia
13.
Cir Esp ; 79(4): 237-40, 2006 Apr.
Artigo em Espanhol | MEDLINE | ID: mdl-16753104

RESUMO

INTRODUCTION: Adrenal incidentalomas are unsuspected, clinically silent adrenal lesions discovered incidentally by imaging tests performed a priori for problems unrelated to the adrenal glands. The aim of this study was to present a series of incidentalomas and review the diagnostic process and treatment techniques. PATIENTS AND METHOD: Of a series of 63 patients studied and/or treated for adrenal disease, there were 34 patients with adrenal incidentalomas. The mean age of the patients with incidentaloma was 50.6 years. There were 23 women (67.6%) and 11 men (32.3%). All patients underwent hormonal investigations to rule out hyperfunction. Imaging techniques consisted of ultrasonography, computed tomography (CT) and/or magnetic resonance imaging (MRI) to determine the size and characteristics of the lesion. In patients with an indication for surgery, the lateral transabdominal approach was used in laparoscopic surgery and the anterior approach was used in open surgery. RESULTS: Of the 34 patients, 23 patients (67.6%) (18 women and 5 men) with a mean age of 50.9 years underwent surgery. The mean tumoral diameter was 10.18 cm. Incidentalomas were located in the right adrenal gland in 16 patients and in the left adrenal gland in 7 patients. Laparoscopic surgery was performed in 9 patients (39.1%) and open surgery was performed in the remaining 14 (60.8%). The mean length of hospital stay was 8.6 days in open surgery and 4 days in laparoscopic surgery. Complications consisted of pneumonia in 2 patients (8.6%). There was no intraoperative mortality in the series. Surgery was not performed in 11 patients (32.3%) (5 women and 6 men) with a mean age of 56.3 years. The mean size of the tumor in these patients, identified by CT, was 2.5 cm. These patients underwent subsequent monitoring with a mean follow-up of 32 months. CONCLUSIONS: Due to the high resolution of ultrasonography, CT and MRI, as well as the greater number of radiological investigations performed, identification of adrenal incidentalomas has increased. In our series these tumors represented 53.9% of adrenal disease.


Assuntos
Neoplasias das Glândulas Suprarrenais/diagnóstico , Neoplasias das Glândulas Suprarrenais/cirurgia , Achados Incidentais , Algoritmos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
14.
Am J Med Qual ; 20(5): 268-76, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-16221835

RESUMO

Primary spontaneous pneumothorax is a common abnormality for which there is a variety of treatments. This study presents the results of a year's evaluation of a clinical pathway (CP). A series of 34 patients treated during 1 year before CP development was analyzed to identify the weak points. To address these weak points, the CP includes associated protocols. In the CP, 31 patients were evaluated during 1 year; the results were compared with those of the pre-CP series. The mean length of stay of the pre-CP patients and the CP patients was 7.3 days and 5.0 days, respectively. The number of radiographs fell from 4.3 to 3.2. The rate of complications and readmissions is similar in both groups. The mean cost per process dropped from 1863 [UNKNOWN] to 1168 [UNKNOWN]. The CP for pneumothorax successfully manages to reduce both the variability in care patterns and hospital costs, justifying the work involved in its development and implementation.


Assuntos
Procedimentos Clínicos , Pneumotórax/terapia , Centro Cirúrgico Hospitalar/organização & administração , Humanos , Pneumotórax/diagnóstico , Espanha
15.
Am Surg ; 71(1): 40-5, 2005 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-15757055

RESUMO

Clinical pathways are comprehensive systematized patient care plans for specific procedures. The clinical pathway for laparoscopic cholecystectomy was implemented in our department in March 2002. The aim of this study is to evaluate the clinical pathway for this procedure 1 year after implementation. A study was conducted on all the patients included in the clinical pathway since its implementation. The assessment criteria include degree of compliance, indicators of clinical care effectiveness, financial impact, and survey-based indicators of satisfaction. The results are compared to a series of patients undergoing surgery the year prior to implementation of the clinical pathway. As our hospital has a system of cost management, we analyzed the mean cost per procedure before and after clinical pathway implementation. Evaluation was made of a series of 160 consecutive patients who underwent surgery during the period 1 year prior to development of the clinical pathway and met the accepted inclusion criteria. The mean length of hospital stay was 3.27 days, and the mean cost per procedure before pathway implementation was 2149 (+/-768) euros. One year after implementation of the pathway, 140 patients were included (i.e., an inclusion rate of 100%). The mean length of hospital stay of the patients included in the clinical pathway was 2.2 days. The degree of compliance with stays was 66.7 per cent. The most frequent reasons for noncompliance were staff-dependent, followed by patient-dependent causes (oral intolerance, pain, etc.). The mean cost in the series of patients included in the clinical pathway was 1845 (+/-618) euros. Laparoscopic cholecystectomy is an ideal procedure for commencing the systemization of clinical pathways. Results show that it has significantly reduced the length of hospital stay and mean cost per procedure with no increased morbidity and with a high degree of patient satisfaction.


Assuntos
Colecistectomia Laparoscópica , Procedimentos Clínicos/organização & administração , Colecistectomia Laparoscópica/economia , Colecistectomia Laparoscópica/normas , Análise Custo-Benefício , Estudos de Avaliação como Assunto , Custos de Cuidados de Saúde/estatística & dados numéricos , Implementação de Plano de Saúde/economia , Humanos , Tempo de Internação/economia , Tempo de Internação/estatística & dados numéricos , Cooperação do Paciente/estatística & dados numéricos , Satisfação do Paciente/estatística & dados numéricos , Estudos Retrospectivos
16.
Cir Esp ; 77(6): 343-50, 2005 Jun.
Artigo em Espanhol | MEDLINE | ID: mdl-16420948

RESUMO

INTRODUCTION: Primary spontaneous pneumothorax (PSP) is a common entity. Treatment varies widely across different health systems and even among doctors in the same department. In our general surgery department 30 to 40 patients with PSP are treated each year and there is significant variability in care delivery. This prompted the development of a clinical pathway (CP) based on analysis of the process, a search for studies with the greatest scientific evidence and professional consensus. The aim of this study was to present the results after 1 year's evaluation of the CP. PATIENTS AND METHOD: A series of 34 patients treated in the year prior to the design of the pathway was analyzed to identify weak points and areas for improvement. To address these weak points the CP included associated protocols and principal documents. Thirty-one patients included in the CP were treated over 1 year and the results were compared with those of the pre-pathway series. RESULTS: The mean length of stay in the pre-pathway patients was 7.3 days compared with 5.0 days in the pathway patients. The number of radiographs also fell from 4.3 to 3.2. The rate of complications and re-admissions was similar in both groups. The mean cost per process decreased from 1863 Euro to 1168 Euro. CONCLUSION: The CP for pneumothorax reduced both variability in professional care patterns and hospital costs, justifying the work involved in its development and implementation.


Assuntos
Pneumotórax/cirurgia , Centro Cirúrgico Hospitalar , Adulto , Progressão da Doença , Feminino , Humanos , Tempo de Internação , Masculino , Pneumotórax/reabilitação
17.
Cancer Gene Ther ; 10(2): 152-60, 2003 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-12536204

RESUMO

Cancer gene therapy based on the use of suicide genes, such as the thymidine kinase gene, is not producing satisfactory results. Several approaches have been delineated to enhance the therapeutic responses, including augmentation of the bystander effect, the combination of the herpes simplex virus thymidine kinase-ganciclovir (HSVTK-GCV) system into replication competent adenoviruses and others. Moreover, because usually less than 20% of human malignant cells are in S-phase, the HSVTK-GCV system is not as efficient as expected. To increase the cytotoxic effects of the HSVTK-GCV system, we hypothesized that concomitant expression of E1a protein, which drives cells to proliferation and S-phase, could increase the effects of the HSVTK-GCV system. Several retroviruses were constructed carrying bicistronic sequences of TK and E1a 12S genes under the control of the CMV promoter. The constructions were tested in murine (NIH-3T3, MSC11A5) and human cells (IMR90, HeLa, MDA-MB435). A clear increase of the HSVTK-GCV system killing effect in nonconfluent cells was observed in the cells studied, especially in NIH-3T3, MSC11A5, IMR90, and MDA-MB435 expressing cells. In confluence, the NIH3T3 and IMR90 E1a-TK-expressing cells were also very sensitive and most malignant E1a-TK-expressing cells showed an irreversible G2-M cell cycle arrest. Moreover, the concomitant expression of adenovirus E1a and the HSVTK-GCV system increased the sensitivity to anticancer agents such as cisplatin. These results show that adenovirus E1a protein expression clearly enhances the cytotoxic effects of the HSVTK-GCV system and the response to treatment with cisplatin.


Assuntos
Proteínas E1A de Adenovirus/farmacologia , Antibióticos Antineoplásicos/farmacologia , Ganciclovir/farmacologia , Terapia Genética/métodos , Simplexvirus/enzimologia , Timidina Quinase/farmacologia , Células 3T3/efeitos dos fármacos , Células 3T3/patologia , Proteínas E1A de Adenovirus/genética , Animais , Ciclo Celular/genética , Cisplatino/farmacologia , Ciclina B/metabolismo , Ciclina B1 , Sinergismo Farmacológico , Engenharia Genética/métodos , Células HeLa/efeitos dos fármacos , Células HeLa/patologia , Humanos , Camundongos , Retroviridae/genética , Simplexvirus/genética , Timidina Quinase/biossíntese , Timidina Quinase/genética , Células Tumorais Cultivadas
18.
Oncogene ; 21(46): 7131-6, 2002 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-12370836

RESUMO

In order to investigate the molecular mechanisms implicated in the induction of chemo sensitivity by adenovirus E1a gene expression, we decided to investigate which signal transduction pathways could be affected by the E1a gene in Human Normal Fibroblast (IMR90). No effect was observed in SAPK pathways (p38MAPK and JNK), but E1a was able to affect the Akt activation mediated by insulin. This result was confirmed by transient transfection experiments performed in Cos-7 cells and also observed in other transformed cell lines such as A431. Furthermore, E1a expression induces a decrease in the basal status of Akt activity. Finally we demonstrated that E1a is able to block the Akt activation mediated by cisplatin and correlates with a sensitive phenotype. In summary, our data demonstrate that specific inhibition of the PI3K/Akt pathway mediates some of the biological properties of E1a such as induction of chemosensitivity.


Assuntos
Proteínas E1A de Adenovirus/genética , Antineoplásicos/farmacologia , Cisplatino/farmacologia , Fosfatidilinositol 3-Quinases/fisiologia , Proteínas Serina-Treonina Quinases , Proteínas Proto-Oncogênicas/fisiologia , Transdução de Sinais , Animais , Células COS , Dano ao DNA , Resistencia a Medicamentos Antineoplásicos , Genes ras , Humanos , Insulina/farmacologia , Fosforilação , Proteínas Proto-Oncogênicas c-akt
19.
Oncol Rep ; 9(4): 897-902, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-12066228

RESUMO

Adenoviruses are ubiquitous viruses related to human mild upper tract respiratory infections. Murine cells are semi-permissive to adenovirus replication, and persistent or abortive infections have been associated with tumorigenic potential. Given that only human lymphoid cells are semi-permissive and abortive infections have been described, we hypothesized that adenovirus could be related to the transformation of human haematopoietic cells. We studied 30 lymphomas, 46 leukemias, 10 reactive lymphadenopathies and 40 normal human spleens. The presence of adenovirus sequences and proteins were studied using PCR, southern-blot, slot-blot, in situ hybridization, immunohistochemistry, and western-blot techniques. By using nested PCR, adenovirus sequences were detectable in about 30% of lymphomas, but in less than 10% of leukemias, reactive lymphadenopathies and normal spleens. In no case were we able to demonstrate adenovirus products by in situ hybridization, immunohistochemistry or western-blot. These results indicate that adenovirus sequences are present in a significant number of human lymphomas, but that the number of positive cells is extremely low and no protein expression could be detected. Therefore, we are unable to conclude that persistent infections of human lymphoid cells by adenovirus is related to a higher risk of developing malignant lymphomas or leukemias.


Assuntos
Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/genética , DNA Viral/análise , Leucemia/virologia , Linfoma/virologia , Proteínas E1A de Adenovirus/genética , Proteínas E1A de Adenovirus/metabolismo , Infecções por Adenovirus Humanos/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Southern Blotting , Estudos de Casos e Controles , Primers do DNA , Feminino , Humanos , Leucemia/patologia , Linfoma/patologia , Masculino , Pessoa de Meia-Idade , Hibridização de Ácido Nucleico , Reação em Cadeia da Polimerase , RNA Viral/análise , Baço/virologia
20.
Rev. méd. Urug ; 14(2): 154-8, ago. 1998. ilus, tab
Artigo em Espanhol | LILACS | ID: lil-246853

RESUMO

La cardiomioplastia dinámica es una técnica quirúrgica utilizada en los pacientes con miocardiopatía dilatada idiopática o cardiopatía isquémica en etapa miocardiopática para tratamiento de la insuficiencia cardíaca. Consiste en envolver el corazón con el músculo dorsal ancho e implementar un cardioestimulador para coordinar la contracción entre el dorsal ancho y el músculo cardíaco. Es de interés conocer las características anatómicas del dorsal ancho en el preoperatorio a fin de determinar su adecuación al procedimiento o proceder a su acondicionamiento previo. A fin de determinar si es posible conocer las caracteristicas de este músculo, previamente a realizar una cirugía de cardiomioplastia dinámica, se evaluaron 30 posibles correlaciones entre variables de mediación incruenta en el cadáver y el dorsal ancho. Las medidas fueron tomadas sobre cadáveres por docentes del Departamento de Anatomía de la Facultad de Medicina de Universidad de la República Oriental del Uruguay. Se encontró que los diámetros subaxilar y subxifoideo y con menor poder de peso del cadáver se correlacionaron con el peso del dorsal ancho. Estos resultados permiten determinar en forma aproximada como será el músculo antes de la cirugía y orientar sobre nuevas investigaciones que deberán ser realizadas


Assuntos
Humanos , Cardiomiopatia Dilatada/cirurgia , Cardiomioplastia , Músculos , Retalhos Cirúrgicos
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