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1.
Int J Gynecol Cancer ; 27(3): 473-478, 2017 03.
Artigo em Inglês | MEDLINE | ID: mdl-28187097

RESUMO

OBJECTIVES: The aims of the study were to evaluate agreement between preoperative endometrial samples and surgical specimens in endometrial carcinoma and to correlate this agreement with sample and patient characteristics. METHODS: Patients who received primary surgical treatment for endometrial carcinoma at a tertiary care center and had undergone preoperative endometrial sampling were included. Medical records were reviewed to collect information from pathology reports and data on patient characteristics. RESULTS: The study sample comprised 166 patients (mean age, 64.6 years). The histological results of the biopsies were the following: endometrioid cancer (n = 118), nonendometrioid tumor (n = 38), and hyperplasia (n = 10). The agreement rates were 93.2% for endometrioid and 68.9% for nonendometrioid tumors, with a κ coefficient of 0.73 for tumor cell type. Tumor International Federation of Gynecology and Obstetrics (FIGO) grade was distributed as follows: 37.1% G1, 35.7% G2, and 27.1% G3, with agreement rates of 61.5%, 56%, and 78.9%, respectively. The overall κ coefficient for FIGO grading was 0.46. Only 1.9% of the tumors originally classified as G1 were upgraded to G3, whereas 16% of G2 lesions were upgraded. There was no significant difference in agreement rates for tumor cell type and FIGO grade in relation to any of the studied variables, except that biopsy specimens weighing more than 3 g had significantly better agreement in FIGO grading (P = 0.040). CONCLUSIONS: Preoperative biopsy has suboptimal accuracy for prediction of characteristics in the definitive surgical specimen. Caution must be taken when using preoperative information to determine extent of surgical resection, due to the risk of understaging. Additional information must be combined with the biopsy data to help in the decision-making process.


Assuntos
Neoplasias do Endométrio/patologia , Neoplasias do Endométrio/cirurgia , Idoso , Biópsia/métodos , Carcinoma Endometrioide/patologia , Carcinoma Endometrioide/cirurgia , Feminino , Humanos , Pessoa de Meia-Idade , Gradação de Tumores , Cuidados Pré-Operatórios/métodos , Estudos Retrospectivos
2.
Int J Gynecol Cancer ; 27(8): 1747-1752, 2017 10.
Artigo em Inglês | MEDLINE | ID: mdl-28937445

RESUMO

OBJECTIVE: The aim of this study was to evaluate the role of follow-up tests and examinations in diagnosing symptomatic and asymptomatic relapses after treatment for cervical cancer. METHODS: Data were collected from medical records for all patients diagnosed as having cervical cancer from January 1985 to June 2010. The significance level was P < 0.005. RESULTS: Sixty-four (17.8%) of the 358 patients investigated suffered tumor relapse. Thirty-four (53.1%) were symptomatic, and 30 (46.9%) were asymptomatic. Most patients had tumor relapse diagnosed during physical examination, both among the symptomatic patients (50%) and the asymptomatic patients (66.7%) (P = 0.27). Cytopathology was responsible for detecting relapse in only 1 case in each group, corresponding to 2.9% and 3.3%, respectively (P = 0.99). Imaging examinations confirmed 10 relapses (29.4%) among symptomatic patients and 8 cases (26.6%) among asymptomatic patients (P = 0.77). There were no statistically significant differences between the 2 groups or between the different methods of detecting relapses. There was still no association after adjustment for potential confounding factors such as age and type of treatment. CONCLUSIONS: Physical examination was the preeminent method for detecting tumor relapse in this study. None of the other tests or examinations were capable of detecting relapses in both symptomatic and asymptomatic patients. These results highlight the urgent need for prospective studies that compare the efficacy of different follow-up regimens, analyzing factors such as global survival, quality of life, and cost.


Assuntos
Recidiva Local de Neoplasia/diagnóstico , Neoplasias do Colo do Útero/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Feminino , Seguimentos , Humanos , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/diagnóstico por imagem , Recidiva Local de Neoplasia/patologia , Exame Físico/métodos , Recidiva , Estudos Retrospectivos , Neoplasias do Colo do Útero/diagnóstico por imagem , Neoplasias do Colo do Útero/patologia , Adulto Jovem
3.
Biochim Biophys Acta ; 1852(7): 1334-46, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25766108

RESUMO

Quiescin sulfhydryl oxidase 1 (QSOX1) is a flavoenzyme largely present in the extracellular milieu whose physiological functions and substrates are not known. QSOX1 has been implicated in the regulation of tumor cell survival, proliferation and migration, in addition to extracellular matrix (ECM) remodeling. However, data regarding other pathophysiological conditions are still lacking. Arterial injury by balloon catheter is an established model of post-angioplasty restenosis. This technique induces neointima formation due to migration and proliferation of vascular smooth muscle cells (VSMC), followed by ECM synthesis and remodeling. Here, we show that QSOX1 knockdown inhibited VSMC migration and proliferation in vitro. In contrast, QSOX1 overexpression stimulated these processes. While migration could be induced by the incubation of cells with the active recombinant QSOX1, proliferation was induced by addition of the active and also of an inactive mutant QSOX1 protein. The proliferation induced by both recombinants was independent of intracellular hydrogen peroxide and dependent of the MEK/ERK pathway. To recapitulate in vivo VSMC pathophysiology, balloon-induced arterial injury was performed. The expression of QSOX1 in the neointimal layer of balloon-injured rat carotids was high and peaked at 14 days post-injury. In vivo QSOX1 knockdown led to a significant decrease in PCNA expression at day 14 post-injury and a decreased intima/media area ratio at day 21 post-injury, compared with scrambled siRNA transfection. In summary, our findings demonstrate that QSOX1 induces VSMC migration and proliferation in vitro and contributes to neointima thickening in balloon-injured rat carotids.


Assuntos
Movimento Celular , Proliferação de Células , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , Neointima/metabolismo , Tiorredoxinas/metabolismo , Animais , Artérias Carótidas/patologia , Artérias Carótidas/cirurgia , Células Cultivadas , Peróxido de Hidrogênio/metabolismo , Sistema de Sinalização das MAP Quinases , Masculino , Camundongos , Músculo Liso Vascular/citologia , Músculo Liso Vascular/fisiologia , Miócitos de Músculo Liso/fisiologia , Ratos , Ratos Wistar , Tiorredoxinas/genética
4.
Nutr Cancer ; 68(6): 935-42, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27340931

RESUMO

We investigated the effect of fish oil (FO) supplementation, at 4 g/day, on the respiratory performance and blood lipid profile of 32 patients with breast cancer at the beginning of chemotherapy. They were randomized into two groups: control (C) and FO supplemented (S). Both groups underwent three respiratory evaluations and blood harvest (before chemotherapy-Day 0, and 30 and 60 days after supplementation). The S group showed a significant increase in the maximal inspiratory and expiratory pressure (P ≤ 0.05 vs. Day 0) and in the maximum voluntary ventilation (P ≤ 0.05). In the treadmill 6-min-walk test, the S group had a significant increase in the walked distance (P ≤ 0.05). Blood lactate concentration was significantly lower in the S group after 60 days, at rest, when compared to C (P ≤ 0.05). Plasma high-density lipoprotein (HDL) cholesterol concentration remained the same after 60 days of supplementation, while in the C group, it decreased significantly (P ≤ 0.05 Day 0 vs. Day 60). Triacylglycerol (TAG) plasma concentration in the S group was lower when compared to the C group (P ≤ 0.05 Day 60S vs. Day 60). Supplementation with FO caused improvement in the respiratory muscle strength and endurance, ameliorated functional performance, and kept TAG, HDL cholesterol, and lactate plasma concentration at normal levels.


Assuntos
Antineoplásicos/efeitos adversos , Neoplasias da Mama/tratamento farmacológico , Suplementos Nutricionais , Óleos de Peixe/uso terapêutico , Pulmão/efeitos dos fármacos , Resistência Física/efeitos dos fármacos , Insuficiência Respiratória/prevenção & controle , Adulto , Antineoplásicos/uso terapêutico , Brasil , Neoplasias da Mama/sangue , Neoplasias da Mama/dietoterapia , Neoplasias da Mama/cirurgia , Quimioterapia Adjuvante/efeitos adversos , HDL-Colesterol/sangue , Suplementos Nutricionais/efeitos adversos , Teste de Esforço , Feminino , Óleos de Peixe/efeitos adversos , Humanos , Ácido Láctico/sangue , Pulmão/fisiopatologia , Pessoa de Meia-Idade , Força Muscular/efeitos dos fármacos , Cuidados Pós-Operatórios , Complicações Pós-Operatórias/induzido quimicamente , Complicações Pós-Operatórias/fisiopatologia , Complicações Pós-Operatórias/prevenção & controle , Reprodutibilidade dos Testes , Insuficiência Respiratória/induzido quimicamente , Insuficiência Respiratória/etiologia , Insuficiência Respiratória/fisiopatologia , Músculos Respiratórios/efeitos dos fármacos , Músculos Respiratórios/fisiopatologia , Triglicerídeos/sangue
5.
Arch Gynecol Obstet ; 288(1): 183-9, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23400355

RESUMO

PURPOSE: This study aims to evaluate and to compare the performance of cervical digital photography (CDP) to the visual inspection with acetic acid (VIA) and visual inspection with Lugol's iodine (VILI) methods for screening the uterine cervix cancer and its precursor lesions in developing countries. METHODS: A cross-sectional study was performed in Brazil. 176 women were evaluated by VIA, VILI, CDP with acetic acid and CDP with Lugol's iodine. Kappa statistic was used to estimate the interobserver and intermethod agreement. Sensitivity, specificity and diagnostic accuracy of the four methods (VIA, VILI, CDP with acetic acid, CDP with Lugol's iodine) was calculated. RESULTS: Interobserver agreement for CDP with acetic acid was K = 0.441 and for CDP with Lugol's iodine was K = 0.533; intermethod agreement of VIA and CDP with acetic acid, K = 0.559; and of VILI and CDP with Lugol's iodine, K = 0.507. Sensitivity and specificity of CDP with acetic acid were 84.00 and 95.83 %, and of CDP with Lugol's iodine were 88.00 and 97.26 %, respectively. The diagnostic accuracy of CDP with acetic acid and CDP with Lugol's iodine was 92.78 and 94.90 %, respectively. CONCLUSION: This was the first study to assess the CDP with Lugol's iodine performance, which had similar performance to the CDP with acetic acid. CDP is considered a promising method for screening the uterine cervix cancer and its precursor lesions in developing countries.


Assuntos
Adenocarcinoma/patologia , Carcinoma de Células Escamosas/patologia , Países em Desenvolvimento , Detecção Precoce de Câncer/métodos , Fotografação , Displasia do Colo do Útero/patologia , Neoplasias do Colo do Útero/patologia , Ácido Acético , Adulto , Brasil , Colo do Útero/patologia , Corantes , Estudos Transversais , Feminino , Humanos , Indicadores e Reagentes , Iodetos , Pessoa de Meia-Idade , Variações Dependentes do Observador , Valor Preditivo dos Testes
6.
J Surg Case Rep ; 2021(8): rjab342, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34408839

RESUMO

Complex reconstructions of the abdominal wall, necessary after resection of neoplasms, infection or trauma, are a challenge for the surgical team. Although ovarian carcinoma is commonly presented with peritoneal carcinomatosis and invasion of adjacent organs, it rarely can invade the abdominal wall. Invasion of the abdominal wall was documented on ultrasound and abdominal computed tomography. Surgery was discussed and performed in a multidisciplinary team and consisted of wide en bloc excision and reconstruction with open intraperitoneal onlay mesh with inorganic polypropylene-coated mesh (Bard/BD Sepramesh), a midweight macroporous mesh and abdominoplasty. Postoperative course was uneventful and the patient showed good evolution 1 year after the procedure. Our report highlights the main objectives in complex reconstructions, the importance of a multidisciplinary team and discusses the characteristics that the mesh must have in order to achieve the desired goal.

7.
Artigo em Inglês | MEDLINE | ID: mdl-33249348

RESUMO

INTRODUCTION: Fish oil (FO) has an anti-inflammatory and pro-resolution activity and it has been used to restore physiological disturbances on inflammatory conditions. Here, we investigate whether FO supplementation could, acutely, prevent or restore inflammatory damages on experimental colitis. METHODS: Wistar rats orally received 2 g.kg-1.day-1 of FO for 30 days before induction of experimental colitis. Specimens were collected on the 2nd and 7th days after colitis-induction and intestinal mucus, inflammatory activity and colon integrity were determined. RESULTS: Experimental colitis did cause colon disruption and FO, acutely, did not prevent the loss of intestinal and fecal mucus, neither the increase of inflammatory activity and intestinal permeability. On the 7th day of colitis, FO soften the perturbations of experimental colitis, increasing histological and fecal mucus and, also decreased inflammatory activity, but this was not accompanied by intestinal permeability. CONCLUSION: FO did not protect, acutely, intestinal damages from experimental colitis, but at long run promotes higher intestinal recovery.


Assuntos
Colite/tratamento farmacológico , Colo/metabolismo , Óleos de Peixe/farmacologia , Animais , Colite/induzido quimicamente , Colite/metabolismo , Colite/patologia , Colo/patologia , Modelos Animais de Doenças , Masculino , Ratos , Ratos Wistar
8.
J Ethnopharmacol ; 255: 112722, 2020 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-32114165

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Euphorbia tirucalli L. is an African plant that grows well in Brazil. Individuals diagnosed with cancer frequently consume latex from E. tirucalli, dissolved in drinking water. In vitro studies confirm the antitumor potential of E. tirucalli latex, but in vivo evaluations are scarce. AIM OF THE STUDY: To evaluate the effect of intake of an aqueous solution of E. tirucalli latex on tumor growth, cachexia, and immune response in Walker 256 tumor-bearing rats. MATERIALS AND METHODS: Latex from E. tirucalli was collected and analyzed by LC-MS. Sixty male Wistar rats (age, 90 days) were randomly divided into four groups: C, control group (without tumor); W, Walker 256 tumor-bearing group; SW1, W animals but treated with 25 µL latex/mL water; and SW2, W animals but treated with 50 µL latex/mL water. Animals received 1 mL of latex solution once a day by gavage. After 15 d, animals were euthanized, tumor mass was determined, and glucose and triacylglycerol serum levels were measured by using commercial kits. Change in the body weight during tumor development was calculated, and proliferation capacity of tumor cells was assessed by the Alamar Blue assay. Phagocytosis and superoxide anion production by peritoneal macrophages and circulating neutrophils were analyzed by enzymatic and colorimetric assays. Data are analyzed by one-way ANOVA followed by Tukey's post-hoc test, with the significance level set at 5%. RESULTS: The analysis of the latex revealed the presence of triterpenes. The ingestion of the latex aqueous solution promoted 40% and 60% reduction of the tumor mass in SW1 and SW2 groups, respectively (p < 0.05). The proliferative capacity of tumor cells from SW2 group was 76% lower than that of cells from W group (p < 0.0001). Animals treated with latex gained, on average, 20 g (SW1) and 8 g (SW2) weight. Glucose and triacylglycerol serum levels in SW1 and SW2 animals were similar to those in C group rats. Peritoneal macrophages and blood neutrophils from SW1 and SW2 animals produced 30-40% less superoxide anions than those from W group animals (p < 0.05), but neutrophils from SW2 group showed an increased phagocytic capacity (20%, vs. W group). CONCLUSIONS: E. tirucalli latex, administered orally for 15 d, efficiently reduced tumor growth and cachexia in Walker 256 tumor-bearing rats. Decreased tumor cell proliferative capacity was one of the mechanisms involved in this effect. Further, the data suggest immunomodulatory properties of E. tirucalli latex. The results agree with folk data on the antitumor effect of latex ingestion, indicating that it may be useful as an adjunct in the treatment of cancer patients. For this, further in vivo studies in animal and human models need to be conducted.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Caquexia/prevenção & controle , Carcinoma 256 de Walker/tratamento farmacológico , Euphorbia , Látex/farmacologia , Extratos Vegetais/farmacologia , Animais , Antineoplásicos Fitogênicos/isolamento & purificação , Biomarcadores/sangue , Glicemia/efeitos dos fármacos , Glicemia/metabolismo , Caquexia/sangue , Caquexia/imunologia , Caquexia/fisiopatologia , Carcinoma 256 de Walker/patologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Euphorbia/química , Látex/isolamento & purificação , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Masculino , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Extratos Vegetais/isolamento & purificação , Ratos Wistar , Triglicerídeos/sangue , Carga Tumoral/efeitos dos fármacos , Redução de Peso/efeitos dos fármacos
9.
Biochim Biophys Acta ; 1780(2): 167-78, 2008 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18082635

RESUMO

Brown spider bites are associated with lesions including dermonecrosis, gravitational spreading and a massive inflammatory response, along with systemic problems that may include hematological disturbances and renal failure. The mechanisms by which the venom exerts its noxious effects are currently under investigation. It is known that the venom contains a major toxin (dermonecrotic toxin, biochemically a phospholipase D) that can experimentally induce dermonecrosis, inflammatory response, animal mortality and platelet aggregation. Herein, we describe cloning, heterologous expression, purification and functionality of a novel isoform of the 33 kDa dermonecrotic toxin. Circular dichroism analysis evidenced correct folding for the toxin. The recombinant toxin was recognized by whole venom serum antibodies and by a specific antibody to a previously described dermonecrotic toxin. The identified toxin was found to display phospholipase activity and dermonecrotic properties. Additionally, the toxin caused a massive inflammatory response in rabbit skin dermis, evoked platelet aggregation, increased vascular permeability, caused edema and death in mice. These characteristics in combination with functional studies for other dermonecrotic toxins illustrate that a family of dermonecrotic toxins exists, and includes a novel member with high activity that may be useful for future structural and functional studies.


Assuntos
Derme/efeitos dos fármacos , Fosfolipase D/química , Fosfolipase D/toxicidade , Venenos de Aranha/química , Venenos de Aranha/enzimologia , Venenos de Aranha/toxicidade , Sequência de Aminoácidos , Animais , Permeabilidade Capilar/efeitos dos fármacos , Clonagem Molecular , DNA Complementar/genética , Derme/patologia , Edema/induzido quimicamente , Camundongos , Dados de Sequência Molecular , Necrose/induzido quimicamente , Fosfolipase D/genética , Diester Fosfórico Hidrolases/genética , Diester Fosfórico Hidrolases/toxicidade , Filogenia , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/toxicidade , Coelhos , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/toxicidade , Venenos de Aranha/genética , Aranhas/enzimologia
10.
Pathol Oncol Res ; 14(1): 23-30, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18398703

RESUMO

The objective of this study was to verify the frequency of p53 and BCL-2 immunohistochemical expression in patients with endometrial carcinoma and to correlate it with histological factors (histological type, tumor grade, depth of myometrial invasion, lymph node involvement and surgical staging) and survival. Forty-eight patients with endometrial carcinoma who were submitted to primary surgical treatment were assessed. p53 and BCL-2 immunohistochemical expression was determined using paraffin blocks containing the tumor area. p53 and BCL-2 expression was detected in 39.6% and 58.3% of the tumors, respectively. No significant difference was found regarding the frequency of p53 expression when analyzing histological type (33.3% in endometrioid tumors, 58.3% in non-endometrioid tumors; p = 0.176), depth of myometrial invasion (p = 0.632) and surgical staging (I-11.1%, II-66.7%, III-57.1%; p = 0.061). p53 expression was significantly more frequent in undifferentiated tumors (p = 0.007) and in those showing lymph node involvement (p = 0.030). Univariate analysis showed a positive association with death (RR, 3.358; CI, 1.386-8.134; p = 0.005) and short-term survival. The present study did not reveal any correlation between BCL-2 expression and histopathologic markers or survival. In conclusion, this study showed that p53 expression is directly correlated with undifferentiated tumors, lymph-node involvement and risk of death. On the other hand, BCL-2 expression was not correlated with any known histological factors.


Assuntos
Biomarcadores Tumorais/análise , Carcinoma Endometrioide/diagnóstico , Neoplasias do Endométrio/diagnóstico , Proteínas Proto-Oncogênicas c-bcl-2/análise , Proteína Supressora de Tumor p53/análise , Adulto , Idoso , Idoso de 80 Anos ou mais , Análise de Variância , Carcinoma Endometrioide/metabolismo , Carcinoma Endometrioide/mortalidade , Carcinoma Endometrioide/patologia , Distribuição de Qui-Quadrado , Neoplasias do Endométrio/metabolismo , Neoplasias do Endométrio/mortalidade , Neoplasias do Endométrio/patologia , Feminino , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Metástase Linfática , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico
11.
Biochem J ; 406(2): 355-63, 2007 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-17535156

RESUMO

Injuries caused by brown spiders (Loxosceles genus) are associated with dermonecrotic lesions with gravitational spreading and systemic manifestations. The venom has a complex composition containing many different toxins, of which metalloproteases have been described in many different species of this genus. These toxins may degrade extracellular matrix constituents acting as a spreading factor. By using a cDNA library from an Loxosceles intermedia venom gland, we cloned and expressed a 900 bp cDNA, which encoded a signal peptide and a propeptide, which corresponded to a 30 kDa metalloprotease, now named LALP (Loxosceles astacin-like protease). Recombinant LALP was refolded and used to produce a polyclonal antiserum, which showed cross-reactivity with a 29 kDa native venom protein. CD analysis provided evidence that the recombinant LALP toxin was folded correctly, was still in a native conformation and had not aggregated. LALP addition to endothelial cell cultures resulted in de-adhesion of the cells, and also in the degradation of fibronectin and fibrinogen (this could be inhibited by the presence of the bivalent chelator 1,10-phenanthroline) and of gelatin in vitro. Sequence comparison (nucleotide and deduced amino acid), phylogenetic analysis and analysis of the functional recombinant toxin revealed that LALP is related in both structure and function to the astacin family of metalloproteases. This suggests that an astacin-like toxin is present in a animal venom secretion and indicates that recombinant LALP will be a useful tool for future structural and functional studies on venom and the astacin family.


Assuntos
Expressão Gênica , Metaloendopeptidases/química , Metaloendopeptidases/metabolismo , Venenos de Aranha/química , Venenos de Aranha/enzimologia , Aranhas/química , Aranhas/enzimologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Dicroísmo Circular , Clonagem Molecular , DNA Complementar/genética , Células Endoteliais/efeitos dos fármacos , Fibrinogênio/metabolismo , Fibronectinas/metabolismo , Gelatina/metabolismo , Humanos , Metaloendopeptidases/genética , Metaloendopeptidases/toxicidade , Dados de Sequência Molecular , Filogenia , Estrutura Secundária de Proteína , Coelhos , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Venenos de Aranha/genética , Venenos de Aranha/toxicidade , Aranhas/classificação , Aranhas/genética
12.
Biochimie ; 89(3): 289-300, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17296256

RESUMO

Loxoscelism (the condition produced by the bite of brown spiders) has been reported worldwide, but especially in warmer regions. Clinical manifestations include skin necrosis with gravitational spreading while systemic loxoscelism may include renal failure, hemolysis and thrombocytopenia. The venom contains several toxins, of which the best biochemically and biologically studied is the dermonecrotic toxin, a phospholipase-D. Purified toxin induces cutaneous and systemic loxoscelism, especially necrotic lesions, hematological disturbances and renal failure. Herein, we describe cloning, heterologous expression and purification of two novel dermonecrotic toxins: LiRecDT4 and LiRecDT5. The recombinant proteins stably expressed in Escherichia coli cells were purified from culture supernatants in a single step using Ni(2+)-chelating chromatography producing soluble proteins of 34 kDa (LiRecDT4) and 37 kDa (LiRecDT5). Circular dichroism analysis evidenced correctly folding for toxins but differences in secondary structures. Both proteins were recognized by whole venom serum antibodies and by a specific antibody to dermonecrotic toxin. Also, recombinant toxins with phospholipase activity induced experimental skin lesions and caused a massive inflammatory response in rabbit skin dermis. Nevertheless, toxins displayed different effects upon platelet aggregation, increase in vascular permeability and not caused death in mice. These characteristics in combination with functional studies illustrates that a family of dermonecrotic toxins exists, and includes two novel members that are useful for future structural and functional studies. They will also be useful in biotechnological ends, for example, as inflammatory and platelet aggregating studies, as antigens for serum therapy source and for lipids biochemical research.


Assuntos
Venenos de Aranha/genética , Venenos de Aranha/metabolismo , Aranhas/genética , Toxinas Biológicas/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Permeabilidade Capilar/efeitos dos fármacos , Dicroísmo Circular , Clonagem Molecular , DNA Complementar/química , DNA Complementar/genética , Eletroforese em Gel de Poliacrilamida , Camundongos , Dados de Sequência Molecular , Fosfolipases/genética , Fosfolipases/metabolismo , Filogenia , Agregação Plaquetária/efeitos dos fármacos , Coelhos , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/toxicidade , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Pele/efeitos dos fármacos , Pele/patologia , Aranhas/metabolismo , Toxinas Biológicas/metabolismo , Toxinas Biológicas/toxicidade
13.
Biochimie ; 88(9): 1241-53, 2006 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16581177

RESUMO

Brown spider (Genus Loxosceles) bites are normally associated with necrotic skin degeneration, gravitational spreading, massive inflammatory response at injured region, platelet aggregation causing thrombocytopenia and renal disturbances. Brown spider venom has a complex composition containing many different toxins, of which a well-studied component is the dermonecrotic toxin. This toxin alone may produce necrotic lesions, inflammatory response and platelet aggregation. Biochemically, dermonecrotic toxin belongs to a family of toxins with 30-35 kDa characterized as sphingomyelinase-D. Here, employing a cDNA library of Loxosceles intermedia venom gland, we cloned and expressed two recombinant isoforms of the dermonecrotic toxin LiRecDT2 (1062 bp cDNA) and LiRecDT3 (1007 bp cDNA) that encode for signal peptides and complete mature proteins. Phylogenetic tree analysis revealed a structural relationship for these toxins compared to other members of family. Recombinant molecules were expressed as N-terminal His-tag fusion proteins in Escherichia coli and were purified to homogeneity from cell lysates by Ni(2+) chelating chromatography, resulting in proteins of 33.8 kDa for LiRecDT2 and 34.0 kDa for LiRecDT3. Additional evidence for related toxins containing sequence/epitopes identity comes from antigenic cross-reactivity using antibodies against crude venom toxins and antibodies raised with a purified dermonecrotic toxin. Recombinant toxins showed differential functionality in rabbits: LiRecDT2 caused a macroscopic lesion with gravitational spreading upon intradermal injection, while LiRecDT3 evoked transient swelling and erythema upon injection site. Light microscopic analysis of skin biopsies revealed edema, a collection of inflammatory cells in and around blood vessels and a proteinaceous network at the dermis. Moreover, differential functionality for recombinant toxins was also demonstrated by a high sphingomyelinase activity for LiRecDT2 and low activity for LiRecDT3 as well as greater in vitro platelet aggregation and blood vessel permeability induced by LiRecDT2 and residual activity for LiRecDT3. Cloning and expression of two recombinant dermonecrotic toxins demonstrate an intraspecific family of homologous toxins that act in synergism for deleterious activities of the venom and open possibilities for biotechnological applications for recombinant toxins as research tools for understanding the inflammatory response, vascular integrity and platelet aggregation modulators.


Assuntos
Diester Fosfórico Hidrolases/química , Diester Fosfórico Hidrolases/genética , Venenos de Aranha/química , Venenos de Aranha/genética , Aranhas/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Camundongos , Dados de Sequência Molecular , Diester Fosfórico Hidrolases/farmacologia , Filogenia , Agregação Plaquetária/efeitos dos fármacos , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/farmacologia , Coelhos , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Alinhamento de Sequência , Esfingomielina Fosfodiesterase/metabolismo , Venenos de Aranha/farmacologia , Aranhas/química
14.
Braz J Microbiol ; 46(3): 753-7, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26413057

RESUMO

Quinolones and fluoroquinolones are widely used to treat uropathogenic Escherichia coli infections. Bacterial resistance to these antimicrobials primarily involves mutations in gyrA and parC genes. To date, no studies have examined the potential relationship between biochemical characteristics and quinolone resistance in uropathogenic E. coli strains. The present work analyzed the quinolone sensitivity and biochemical activities of fifty-eight lactose-negative uropathogenic E. coli strains. A high percentage of the isolates (48.3%) was found to be resistant to at least one of the tested quinolones, and DNA sequencing revealed quinolone resistant determining region gyrA and parC mutations in the multi-resistant isolates. Statistical analyses suggested that the lack of ornithine decarboxylase (ODC) activity is correlated with quinolone resistance. Despite the low number of isolates examined, this is the first study correlating these characteristics in lactose-negative E. coli isolates.


Assuntos
Farmacorresistência Bacteriana Múltipla/genética , Infecções por Escherichia coli/tratamento farmacológico , Fluoroquinolonas/uso terapêutico , Lactose/metabolismo , Ácido Nalidíxico/uso terapêutico , Ornitina Descarboxilase/genética , Infecções Urinárias/tratamento farmacológico , Escherichia coli Uropatogênica/genética , Antibacterianos/uso terapêutico , Brasil , DNA Girase/genética , DNA Topoisomerase IV/genética , Descarboxilação/genética , Descarboxilação/fisiologia , Infecções por Escherichia coli/microbiologia , Humanos , Testes de Sensibilidade Microbiana , Ornitina/metabolismo , Infecções Urinárias/microbiologia , Escherichia coli Uropatogênica/efeitos dos fármacos , Escherichia coli Uropatogênica/enzimologia , Escherichia coli Uropatogênica/isolamento & purificação
15.
Toxicon ; 44(7): 693-709, 2004 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-15500846

RESUMO

Accidents caused by brown spiders (Loxosceles genus) are classically associated with dermonecrotic lesions and systemic manifestations including intravascular haemolysis, disseminated intravascular coagulation and acute renal failure. Systemic reactions occur in a minority of cases, but may be severe in some patients and occasionally fatal. The mechanisms by which Loxosceles venom exerts these noxious effects are currently under investigation. The venom contains several toxins, some of which have been well-characterised biochemically and biologically. The purpose of the present review is to describe some insights into loxoscelism obtained over the last ten years. The biology and epidemiology of the brown spider, the histopathology of envenomation and the immunogenicity of Loxosceles venom are reviewed, as are the clinical features, diagnosis and therapy of brown spider bites. The identification and characterisation of some toxins and the mechanism of induction of local and systemic lesions caused by brown spider venom are also discussed. Finally, the biotechnological application of some venom toxins are covered.


Assuntos
Diester Fosfórico Hidrolases/química , Diester Fosfórico Hidrolases/imunologia , Picada de Aranha/epidemiologia , Picada de Aranha/patologia , Venenos de Aranha/química , Venenos de Aranha/imunologia , Aranhas/química , Animais , Biotecnologia , Brasil/epidemiologia , Diester Fosfórico Hidrolases/toxicidade , Picada de Aranha/diagnóstico , Picada de Aranha/terapia , Venenos de Aranha/toxicidade , Aranhas/fisiologia
16.
Chem Biol Interact ; 206(1): 37-46, 2013 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-23994248

RESUMO

Galectin-3, a ubiquitous member of the galectin family, has been shown to control cellular proliferation, adhesion, migration and apoptosis; thus, it has a role in tumor development and progression. Galectin-3 expression is both up- and down-regulated during melanoma progression. However, conflicting data regarding its roles in tumor biology prompted us to investigate if the presence of galectin-3 influences the response of melanoma cells to a novel metallodrug because metastatic melanoma acquires chemo resistance and is reported to be redox-sensitive. Previously, it was demonstrated that the complex [bis-(2-oxindol-3-yl-imino)-2-(2-aminoethyl) pyridine-N,N'] copper (II) perchlorate, herein referred to as [Cu(isaepy)], induces ROS formation and apoptosis in neuroblastoma cells through mitochondrial uncoupling and the activation of AMPK/p38/p53 signaling. Here, we used a model of vertical growth melanoma (TM1), in which GAL3 expression is lost during tumor progression. When de novo expressed, galectin-3 was found to be ubiquitously present in all subcellular compartments. Our results demonstrate that de novo galectin-3 expression impairs the cellular antioxidant system and renders TM1G3 cells more susceptible than GAL3-null TM1MNG3 cells to [Cu(isaepy)] treatment. This compound, in contrast with the redox inactive [dichloro (2-oxindol-3-yl-imino)-2-(2-aminoethyl) pyridine-N,N'] zinc (II), herein referred to as [Zn(isaepy)], leads to increased intracellular ROS accumulation, increased carbonyl stress, increased mitochondrial depolarization, decreased cell adhesion, increased p38 activation and apoptosis in TM1G3, compared with TM1MNG3. Cell death was shown to be dependent on a hydrogen peroxide-derived species and on the activation of p38. Because mitochondria are a target of both [Cu(isaepy)] and galectin-3, we propose that the presence of galectin-3 in this organelle favors increased ROS production, thereby inducing oxidative cellular damage and apoptotic death. Therefore, [Cu(isaepy)] may be envisaged as a possible anti-melanoma strategy, particularly for melanomas that express galectin-3.


Assuntos
Cobre/farmacologia , Galectina 3/biossíntese , Melanoma/metabolismo , Compostos Organometálicos/farmacologia , Animais , Apoptose/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cobre/química , Relação Dose-Resposta a Droga , Galectina 3/metabolismo , Melanoma/patologia , Camundongos , Compostos Organometálicos/química , Oxirredução , Relação Estrutura-Atividade
17.
Femina ; 43(1)jan.-fev. 2015.
Artigo em Português | LILACS | ID: lil-754438

RESUMO

Este artigo objetivou oferecer uma visão atual do papel dos marcadores gênicos no carcinoma de endométrio. Os principais genes descritos são o TP53, o Bcl-2, o c-erbB2 e o p16. Nos últimos anos, com a ampliação do conhecimento na área de biologia molecular, tem sido sugerido que os marcadores biológicos possam ser tão ou mais importantes do que os fatores prognósticos convencionais.


The main of this study is offer the present situation of genic markers in endometrial carcinoma. The principal genes have been described are TP53, Bcl-2, c-erbB2 and p16. In the last few years, thanks to improvements in molecular biology, some biological markers have been suggested to be as important as or more important than conventional prognostic factors.


Assuntos
Humanos , Feminino , Adulto , Neoplasias do Endométrio , Biomarcadores/análise , /fisiologia , /fisiologia , Histerectomia , Marcadores Genéticos/genética
18.
Clin. biomed. res ; 35(1): 27-34, 2015. tab
Artigo em Português | LILACS | ID: lil-780279

RESUMO

Neste estudo, descreve-se o perfil clínico das pacientes e as características histopatológicas dos carcinomas de endométrio tratados no setor de Oncologia Genital do Hospital de Clínicas de Porto Alegre (HCPA), assim como as formas de tratamento, fatores prognósticos e sobrevida. Métodos: Estudo de coorte histórica incluindo todas as pacientes submetidas a tratamento cirúrgico primário entre 1996 e 2012. Após revisão de prontuários médicos, foram analisadas as variáveis idade, status hormonal, tipo histológico e grau tumoral, invasão miometrial, estadiamento cirúrgico, cirurgia realizada, tratamento complementar e sobrevida. Resultados: Cento e sessenta e quatro pacientes foram incluídas no estudo, com idade média de 64,2 anos (31-95 anos), sendo quase 90% delas pós-menopáusicas. O tempo de seguimento variou de 4 dias a 14,6 anos. O tipo histológico endometrioide foi o mais encontrado (78% dos casos). A histerectomia com salpingo-ooforectomia bilateral com linfadenectomia pélvica foi a cirurgia mais realizada (77,5%). Tratamento complementar foi realizado em 57,9% das pacientes, sendo a radioterapia o tratamento de escolha em 87,4% deles. Ocorreram 36 óbitos (22%) durante o seguimento, com uma sobrevida média global de 125 meses. Em análise bivariada, idade ≥ 65 anos, tipo histológico não endometrioide, tumores pouco diferenciados (G3), invasão miometrial ≥ 50% e metástase linfonodal relacionaram-se significativamente a um menor tempo de sobrevida. Em análise multivariada, a histologia não endometrioide, estádio III, estádio IV e a presença de comprometimento linfonodal foram significativamente associados ao óbito. Conclusão: Os resultados encontrados são compatíveis com a literatura existente e vêm em acréscimo à escassa estatística nacional...


This study describes the clinical profile and the hystopathologic characteristics of endometrial carcinomas from patients treated at the Gynecologic Oncology department of Hospital de Clínicas de Porto Alegre (HCPA), as well as the forms of treatment, prognostic factors, and survival. Methods: Historic cohort study including all patients subjected to primary surgical treatment between 1996 and 2012. After review of the medical records, the variablesage, hormonal status, tumor histologic type and grade, myometrial invasion, surgical staging, performed surgery, complementary treatment, and survival were analyzed. Results: One hundred sixty four patients were included, with a mean age of 64.2 years (31-95 years), of which almost 90% were postmenopausal women. Follow-up time ranged from 4 days to 14.6 years. Endometrioid adenocarcinoma was the most frequently histological type (78% of cases). Hysterectomy with bilateral salpingooophorectomy plus pelvic linfadenectomy was the most frequently performed surgery (77.5%). Adjuvant treatment was held in 57.9% of the patients, with radiotherapy being the treatment of choice in 87.4%. Thirty-six deaths (22%) occurred during followup, with a mean overall survival of 125 months. In the bivariate analysis, age ≥ 65 years, non-endometrioid histology, poorly differentiated tumors (G3), myometrial invasion ≥ 50%, and lymph node metastasis were correlated to lower survival. In the multivariate analysis, non-endometrioid histology, stage III, stage IV and lymph node metastasis were significantly associated with death. Conclusion: The results found are compatible with the existing literature and contribute to the scarce existing national statistics...


Assuntos
Humanos , Feminino , Adenocarcinoma/cirurgia , Adenocarcinoma/mortalidade , Adenocarcinoma/radioterapia , Neoplasias do Endométrio/epidemiologia
19.
Braz. j. microbiol ; 46(3): 753-757, July-Sept. 2015. tab, ilus
Artigo em Inglês | LILACS | ID: lil-755797

RESUMO

Quinolones and fluoroquinolones are widely used to treat uropathogenic Escherichia coli infections. Bacterial resistance to these antimicrobials primarily involves mutations in gyrA and parC genes. To date, no studies have examined the potential relationship between biochemical characteristics and quinolone resistance in uropathogenic E. coli strains. The present work analyzed the quinolone sensitivity and biochemical activities of fifty-eight lactose-negative uropathogenic E. coli strains. A high percentage of the isolates (48.3%) was found to be resistant to at least one of the tested quinolones, and DNA sequencing revealed quinolone resistant determining region gyrA and parC mutations in the multi-resistant isolates. Statistical analyses suggested that the lack of ornithine decarboxylase (ODC) activity is correlated with quinolone resistance. Despite the low number of isolates examined, this is the first study correlating these characteristics in lactose-negative E. coli isolates.

.


Assuntos
Humanos , Farmacorresistência Bacteriana Múltipla/genética , Infecções por Escherichia coli/tratamento farmacológico , Fluoroquinolonas/uso terapêutico , Lactose/metabolismo , Ácido Nalidíxico/uso terapêutico , Ornitina Descarboxilase/genética , Infecções Urinárias/tratamento farmacológico , Escherichia coli Uropatogênica/genética , Antibacterianos/uso terapêutico , Brasil , DNA Girase/genética , DNA Topoisomerase IV/genética , Descarboxilação/genética , Descarboxilação/fisiologia , Infecções por Escherichia coli/microbiologia , Testes de Sensibilidade Microbiana , Ornitina/metabolismo , Infecções Urinárias/microbiologia , Escherichia coli Uropatogênica/efeitos dos fármacos , Escherichia coli Uropatogênica/enzimologia , Escherichia coli Uropatogênica/isolamento & purificação
20.
Int J Exp Pathol ; 83(6): 287-94, 2002 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-12657137

RESUMO

Loxoscelism, the term used to describe envenomation with brown spiders, is characterized by a dermonecrotic lesion at the bite site. In the present investigation we submitted albino rabbits to an acute experimental envenomation protocol using Loxosceles intermedia (brown spider) venom, with in order to determine the pathogenesic features of the lesion induced by this spider, which is the cause of several accidents throughout the world. Rabbits received intradermal injections of the venom and were monitored over the first 4 h, and then at 12 h and 1, 2 and 5 days after envenomation. Histological specimens from 3 rabbits per time point were collected from euthanized animals and processed for histological examination by light microscopy. Major findings observed during the first 4 h were oedema, haemorrhage, degeneration of blood vessel walls, plasma exudation, thrombosis, neutrophil accumulation in and around blood vessels with an intensive diapedesis, a diffuse collection of inflammatory cells (polymorphonuclear leucocytes) in the dermis, and subcutaneous muscular oedema. Over the following hours and up to 5 days after envenomation the changes progressed to massive neutrophil infiltration (with no other leucocytes) into the dermis and even into subcutaneous muscle tissue, destruction of blood vessels, thrombosis, haemorrhage, myonecrosis, and coagulative necrosis on the 5th day.


Assuntos
Músculo Esquelético/patologia , Pele/patologia , Picada de Aranha/patologia , Venenos de Aranha/toxicidade , Animais , Vasos Sanguíneos/patologia , Edema/induzido quimicamente , Edema/patologia , Hemorragia/induzido quimicamente , Injeções Intradérmicas , Músculo Esquelético/imunologia , Necrose , Infiltração de Neutrófilos , Coelhos , Pele/imunologia , Trombose/induzido quimicamente , Fatores de Tempo
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