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1.
Ann Vasc Surg ; 2024 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-38825068

RESUMO

INTRODUCTION: Gupta Perioperative Risk for Myocardial Infarction or Cardiac Arrest (MICA) is a validated self-explanatory score applied in cardiac or non-cardiac surgeries. This study aims to assess the predictive value of the MICA score for cardiovascular events after aortoiliac revascularization. METHODS: This prospective cohort underwent elective aortoiliac revascularization between 2013 and 2021. Patients' demographic, clinical characteristics and outcomes were registered. The patients were divided into two groups according to the MICA score using optimal binning. Survival analysis to test for time-dependent variables and multivariate Cox regression analysis for independent predictors were performed. RESULTS: This study included 130 patients with a median follow-up of 55 months. Preoperative MICA score was ≥ 6.5 in 41 patients. MICA ≥ 6.5 presented a statistically significant association with long-term occurrence of acute heart failure (HR=1.695, 95% CI 1.208-2.379, p=0.002), major adverse cardiovascular events (HR=1.222, 95% CI 1.086-1.376, p<0.001) and all-cause mortality (HR=1.256, 95% CI 1.107-1.425, p<0.001). Multivariable Cox regression confirmed MICA as a significant independent predictor of long-term major adverse cardiovascular events (aHR=1.145 95%CI 1.010-1.298, p=0.034) and all-cause mortality (aHR=1.172 95%CI 1.026-1.339, p=0.020). CONCLUSION: The MICA score is a quick, easy-to-obtain, predictive tool in identifying patients with a higher risk of post-aortoiliac revascularization cardiovascular events, such as acute heart failure, major adverse cardiovascular events and all-cause mortality. Additional research for validation of the MICA score in the context of aortoiliac revascularization and specific interventions are necessary.

2.
Cell Mol Life Sci ; 79(1): 11, 2021 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-34951683

RESUMO

Trichomonas vaginalis is a common sexually transmitted extracellular parasite that adheres to epithelial cells in the human urogenital tract. Extracellular vesicles (EVs) have been described as important players in the pathogenesis of this parasite as they deliver proteins and RNA into host cells and modulate parasite adherence. EVs are heterogeneous membrane vesicles released from virtually all cell types that collectively represent a new dimension of intercellular communication. The Endosomal Sorting Complex Required for Transport (ESCRT) machinery contributes to several key mechanisms in which it reshapes membranes. Based on this, some components of the ESCRT have been implicated in EVs biogenesis in other cells. Here, we demonstrated that VPS32, a member of ESCRTIII complex, contribute to the biogenesis and cargo sorting of extracellular vesicles in the parasite T. vaginalis. Moreover, we observe that parasites overexpressing VPS32 have a striking increase in adherence to host cells compared to control parasites; demonstrating a key role for this protein in mediating host: parasite interactions. These results provide valuable information on the molecular mechanisms involved in extracellular vesicles biogenesis, cargo-sorting, and parasite pathogenesis.


Assuntos
Complexos Endossomais de Distribuição Requeridos para Transporte/metabolismo , Vesículas Extracelulares/metabolismo , Interações Hospedeiro-Parasita , Parasitos/citologia , Trichomonas vaginalis/citologia , Animais , Adesão Celular , Linhagem Celular , Vesículas Extracelulares/ultraestrutura , Humanos , Masculino , Parasitos/metabolismo , Próstata/parasitologia , Espectrometria de Massas em Tandem , Trichomonas vaginalis/metabolismo
3.
Ann Vasc Surg ; 79: 153-161, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34644633

RESUMO

OBJECTIVES: Vascular surgery patients commonly have several comorbidities that cumulatively lead to a frailty status. The cumulative comorbidities disproportionately increase the risk of adverse events and are also associated with worsened long-term prognosis. In recent years, several tools have been elaborated with the objective of quantifying a patient's frailty. One of them is the modified frailty index-5 (mFI-5), a simplified and easy to use index. There is scarce data regarding its value as a prognostic factor in aortoiliac occlusive disease. The aim of this work is to validate mFI-5 as a potential postoperative prognostic indicator in this population. METHODS: From January 2013 to January 2020, 109 patients who underwent elective revascularizations, either endovascular or open surgery, having Trans-Atlantic Inter-Society Consensus II type D aortoiliac lesions in a tertiary and a regional hospital were selected from a prospective vascular registry. Demographic data was collected including diabetes mellitus, chronic heart failure, chronic obstructive pulmonary disease, arterial hypertension requiring medication and functional status. The 30-d and subsequent long-term surveillance outcomes were also collected including major adverse cardiovascular events (MACE), major adverse limb events (MALE) and all-cause mortality were assessed in the 30-d post-procedure and in the subsequent long-term surveillance period. The mFI-5 was applied to this population to evaluate the prognostic impact of this frailty marker on mortality and morbidity. RESULTS: In the long-term follow-up, mFI-5 was significantly associated with MACE (hazard ratio [HR] 2.469; 95% confidence interval [CI]: 1.267-4.811; P = .008) and all-cause mortality (HR 2.585; 95% CI: 1.270-5.260; P = .009). However, there was no significant association with 30-day outcomes. Along with the presence of chronic kidney disease, mFI-5 was the prognostic factor better able of predicting MACE. No prognostic value was found regarding short-term outcomes. CONCLUSION: The mFI-5 index may have a role in predicting long term outcomes, namely MACE and all-cause mortality, in the subset of patients with extensive aortoiliac occlusive disease. Its ease of use can foster its application in risk stratification and contribute for the decision-making process.


Assuntos
Doenças da Aorta/cirurgia , Arteriopatias Oclusivas/cirurgia , Procedimentos Endovasculares , Idoso Fragilizado , Fragilidade/complicações , Artéria Ilíaca/cirurgia , Procedimentos Cirúrgicos Vasculares , Fatores Etários , Idoso , Doenças da Aorta/diagnóstico por imagem , Doenças da Aorta/mortalidade , Arteriopatias Oclusivas/complicações , Arteriopatias Oclusivas/diagnóstico por imagem , Arteriopatias Oclusivas/mortalidade , Comorbidade , Procedimentos Endovasculares/efeitos adversos , Procedimentos Endovasculares/mortalidade , Feminino , Fragilidade/diagnóstico , Fragilidade/mortalidade , Nível de Saúde , Humanos , Artéria Ilíaca/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/etiologia , Complicações Pós-Operatórias/mortalidade , Estudos Prospectivos , Sistema de Registros , Medição de Risco , Fatores de Risco , Fatores de Tempo , Resultado do Tratamento , Procedimentos Cirúrgicos Vasculares/efeitos adversos , Procedimentos Cirúrgicos Vasculares/mortalidade
4.
Ann Vasc Surg ; 81: 216-224, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-34748948

RESUMO

BACKGROUND: Carotid restenosis following carotid endarterectomy (CEA) has a cumulative risk at 5-years up to 32%, which may impact the well-being of patients following CEA. Haematological parameters in the standard complete blood cell count (CBC) are emerging as potential biomarkers, but their application in CEA is scarce. The primary aim of this study was to investigate haematological markers for restenosis following CEA. The secondary aim was to characterize clinical risk factors for restenosis. METHODS: From January 2012 to January 2019, 151 patients who underwent CEA under regional anaesthesia due to carotid stenosis were selected from a prospectively maintained cohort database. Patients were included if a preoperative CBC was available in the 2 weeks preceding CEA. Multivariable analysis was performed alongside propensity score matching (PSM) analysis, using the preoperative CEA parameters, to reduce confounding factors between categories. RESULTS: The study group comprised 28 patients who developed carotid restenosis. The remaining 123 patients without restenosis composed the control group. Mean age of the patients did not differ significantly between groups (70.25 ± 8.05 vs. 70.32 ± 9.61 YO, P = 0.973), neither did gender (male gender 89.3% vs. 78.9%, P = 0.206). Regarding haematological parameters, only MPV remained statistically significant within multivariable analysis (1.855, aOR [1.174-2.931], P = 0.008), a result supported by PSM analysis (2.072, aOR [1.036-4.147], P = 0.042). CONCLUSIONS: MPV was able to predict restenosis 2 years after CEA. Thus, MPV can be incorporated into score calculations to identify patients at greater risk of restenosis, who could benefit from specific monitoring during follow-up. While results are promising, more research is necessary to corroborate them.


Assuntos
Estenose das Carótidas , Endarterectomia das Carótidas , Acidente Vascular Cerebral , Estenose das Carótidas/complicações , Estenose das Carótidas/diagnóstico por imagem , Estenose das Carótidas/cirurgia , Endarterectomia das Carótidas/efeitos adversos , Endarterectomia das Carótidas/métodos , Humanos , Masculino , Volume Plaquetário Médio , Recidiva , Estudos Retrospectivos , Fatores de Risco , Acidente Vascular Cerebral/etiologia , Fatores de Tempo , Resultado do Tratamento
5.
Ann Vasc Surg ; 79: 247-255, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34699941

RESUMO

Contralateral carotid stenosis (clCS) has been described as a perioperative predictor of mortality after carotid endarterectomy (CEA). However, its predictive value on long-term cardiovascular events remains controversial. The study aims to assess the potential role of clCS as a long-term predictor of major adverse cardiovascular events (MACE) in patients who underwent CEA. From January 2012 to July 2020, patients undergoing CEA under regional anesthesia for carotid stenosis in a tertiary care and referral center were eligible from a prospective database, and a post hoc analysis was performed. The primary outcome consisted in the occurrence of long-term MACE. Secondary outcomes included all-cause mortality, stroke, myocardial infarction, acute heart failure, and major adverse limb events. A total of 192 patients were enrolled. With a median 50 months follow-up, chronic kidney disease (CKD) (mean survival time (MST) 51.7 vs. 103.3, P < 0.010) and peripheral artery disease (PAD) (MST 75.1 vs. 90.3, P = 0.001) were associated with decreased survival time. After propensity score matching (PSM), CKD (MST 49.1 vs. 106.0, P = 0.001) and PAD (MST 75.7 vs. 94.0, P = 0.001) maintained this association. On multivariate Cox regression analysis, contralateral stenosis was associated with higher MACE (hazard ratio (HR) = 2.035; 95% CI: 1.113-3.722, P = 0.021 and all-cause mortality (HR = 2.564; 95% CI: 1.276-5,152 P = 0.008). After PSM, only all-cause mortality (HR 2.323; 95% CI: 0.993-5.431, P = 0.052) maintained a significant association with clCS. On multivariable analysis, clCS (aHR 2.367; 95% CI: 1.174-4.771, P = 0.016), age (aHR 1.039, 95% CI: 1.008-1.070), CKD (aHR 2.803; 95% CI: 1.409-5.575, P = 0.003) and PAD (aHR 3.225, 95% CI: 1.695-6.137, P < 0.001) were independently associated with increased all-cause mortality. Contrary to MACE, clCS is a strong predictor of long-term all-cause mortality after CEA. However, MACE risk may compromise CEA benefits by other competitive events. Therefore, further studies are needed to establish the role of clCS on postoperative events and on patients' specific assessments in order to determine the best medical treatment and easy access to surgical intervention.


Assuntos
Estenose das Carótidas/cirurgia , Endarterectomia das Carótidas/efeitos adversos , Complicações Pós-Operatórias/etiologia , Idoso , Estenose das Carótidas/diagnóstico por imagem , Estenose das Carótidas/mortalidade , Endarterectomia das Carótidas/mortalidade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/diagnóstico , Complicações Pós-Operatórias/mortalidade , Valor Preditivo dos Testes , Medição de Risco , Fatores de Risco , Fatores de Tempo , Resultado do Tratamento
6.
Vasa ; 51(2): 93-98, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35171024

RESUMO

Background: Cardiac complications represent the main cause of mortality after non-cardiac surgery and the Revised Cardiac Risk Index (RCRI) was created to estimate the perioperative risk of these events. It considers history of ischaemic heart disease, congestive heart failure, diabetes requiring preoperative insulin, stroke or transient ischaemic attack and renal impairment. We aim to describe the accuracy of the RCRI for predicting perioperative major adverse cardiovascular events (MACE) - a composite of heart failure, ischemic events and all-cause death. Also, the authors aimed to review the score for better prediction of cardiovascular outcomes. Patients and methods: From January 2012 to January 2020, patients who underwent Carotid endarterectomy (CEA) with regional anaesthesia (RA) were selected. RCRI was calculated for each case. Estimated and reported cardiovascular complications were compared using multivariate logistic regression and cox proportional hazards. An alternative and optimized carotid-RCRI (CtRCRI) was obtained. Overall predictive accuracy was assessed and compared by measuring model discrimination. Adjustments for overfitting and evaluation of the new model were performed by bootstrap. Results: 186 patients were selected, of which 80% were male with a mean age of 70.0±9.05 years old. The median follow-up was 50 months, interquartile range 21-69 months. None of the scores were able to predict MACE in the perioperative period. Both were associated with 30-day Clavien-Dindo ≥2 (p=0.022 and p=0.041, respectively). Regarding long-term prognosis, both were able to predict MACE (RCRI: hazard ratio (HR) 3.54 (95% confidence interval [CI] 1.04-11.48) vs. CtRCRI: HR 2.08 (95%CI 1.08-3.98) and all-cause mortality (RCRI: HR 3.33, 95%CI 0.99-11.11 vs. CtRCRI: HR 1.57, 95%CI 1.14-7.04). Conclusions: RCRI and CtRCRI did not predict MACE in the perioperative period but are good predictors of 30-day complications (Clavien-Dindo ≥2). Both RCRI and CtRCRI have good prognostic value as predictors of long-term cardiovascular events.


Assuntos
Endarterectomia das Carótidas , Idoso , Endarterectomia das Carótidas/efeitos adversos , Humanos , Masculino , Pessoa de Meia-Idade , Modelos de Riscos Proporcionais , Estudos Retrospectivos , Medição de Risco , Fatores de Risco
7.
Eur J Vasc Endovasc Surg ; 62(4): 522-531, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34284934

RESUMO

OBJECTIVE: Intra-operative near infrared spectroscopy (NIRS) is a non-invasive tool used to monitor regional cerebral oxygen saturation during carotid endarterectomy (CEA), for which accuracy remains unclear. Therefore, this systematic review and meta-analysis aimed to determine the diagnostic accuracy of NIRS in patients undergoing CEA under regional anaesthesia (RA). DATA SOURCES: MEDLINE, Scopus, and Web of Science were searched for studies that compared NIRS with the "awake test" in patients undergoing CEA under RA. REVIEW METHODS: Bivariable random effects meta-analysis was performed to determine the diagnostic accuracy of NIRS to detect cerebral ischaemia. Meta-regression was performed to explore causes of heterogeneity. Meta-analysis of proportions was also performed to determine the accuracy of NIRS in predicting 30 day stroke. Study quality was evaluated using the QUADAS-2 criteria. RESULTS: Eleven primary studies were included, assessing 1 237 participants. The meta-analysis obtained a partial area under the summary receiver operating characteristic curve for diagnosing brain ischaemia of 0.646, with a summary sensitivity of 72.0% (95% confidence interval [CI] 58.1 - 82.7; I2 = 48.6%) and a specificity of 84.1% (95% CI 78.5-88.4; I2 = 48.6%). In meta-regression analysis, the frequency of hypertension (p = .011) and patients with symptomatic carotid stenosis (p = .031) were significant effect modifiers. Higher frequency of arterial hypertension (z score = -2.15; p = .032) and diabetes (z score = -2.12; p = .034) were associated with lower summary sensitivity, while a higher frequency of symptomatic carotid stenosis (z score = 2.11; p = .035) was associated with higher summary sensitivity. Point estimate sensitivity and specificity for predicting 30 day stroke occurrence were 41% (95% CI 19.5 - 66.6; I2 = 0%) and 81.4% (95% CI 74.4 - 86.9, I2 = 65.6%), respectively. CONCLUSION: The results of this study suggest that NIRS as a cerebral monitoring technique does not have sufficiently high sensitivity or specificity to be used alone in the neurological monitoring of patients undergoing CEA under RA.


Assuntos
Anestesia por Condução , Isquemia Encefálica/diagnóstico , Estenose das Carótidas/cirurgia , Circulação Cerebrovascular , Endarterectomia das Carótidas , Monitorização Intraoperatória , Oxigênio/sangue , Espectroscopia de Luz Próxima ao Infravermelho , Idoso , Anestesia por Condução/efeitos adversos , Biomarcadores/sangue , Isquemia Encefálica/sangue , Isquemia Encefálica/etiologia , Isquemia Encefálica/fisiopatologia , Estenose das Carótidas/sangue , Estenose das Carótidas/diagnóstico , Estenose das Carótidas/fisiopatologia , Endarterectomia das Carótidas/efeitos adversos , Feminino , Humanos , Masculino , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Fatores de Risco , Resultado do Tratamento
8.
Appl Environ Microbiol ; 86(7)2020 03 18.
Artigo em Inglês | MEDLINE | ID: mdl-32005737

RESUMO

The binary (Bin) toxin from Lysinibacillus sphaericus is effective to mosquito larvae, but its utilization is threatened by the development of insect resistance. Bin toxin is composed of the BinB subunit required for binding to midgut receptors and the BinA subunit that causes toxicity after cell internalization, mediated by BinB. Culex quinquefasciatus resistance to this toxin is caused by mutations that prevent expression of Bin toxin receptors in the midgut. Previously, it was shown that the Cyt1Aa toxin from Bacillus thuringiensis subsp. israelensis restores Bin toxicity to Bin-resistant C. quinquefasciatus and to Aedes aegypti larvae, which are naturally devoid of functional Bin receptors. Our goal was to elucidate the mechanism involved in Cyt1Aa synergism with Bin in such larvae. In vivo assays showed that the mixture of Bin toxin, or its BinA subunit, with Cyt1Aa was effective to kill resistant larvae. However, no specific binding interaction between Cyt1Aa and the Bin toxin, or its subunits, was observed. The synergy between Cyt1Aa and Bin toxins is dependent on functional Cyt1Aa, as demonstrated by using the nontoxic Cyt1AaV122E mutant toxin affected in oligomerization and membrane insertion, which was unable to synergize Bin toxicity in resistant larvae. The synergism correlated with the internalization of Bin or BinA into anterior and medium midgut epithelial cells, which occurred only in larvae treated with wild-type Cyt1Aa toxin. This toxin is able to overcome failures in the binding step involving BinB receptor by allowing the internalization of Bin toxin, or its BinA subunit, into the midgut cells.IMPORTANCE One promising management strategy for mosquito control is the utilization of a mixture of L. sphaericus and B. thuringiensis subsp. israelensis insecticidal toxins. From this set, Bin and Cyt1Aa toxins synergize and display toxicity to resistant C. quinquefasciatus and to A. aegypti larvae, whose midgut cells lack Bin toxin receptors. Our data set provides evidence that functional Cyt1Aa is essential for internalization of Bin or its BinA subunit into such cells, but binding interaction between Bin and Cyt1Aa is not observed. Thus, this mechanism contrasts with that for the synergy between Cyt1Aa and the B. thuringiensis subsp. israelensis Cry toxins, where active Cyt1Aa is not necessary but a specific binding between Cry and Cyt1Aa is required. Our study established the initial molecular basis of the synergy between Bin and Cyt1Aa, and these findings enlarge our knowledge of their mode of action, which could help to develop improved strategies to cope with insect resistance.


Assuntos
Aedes/efeitos dos fármacos , Bacillaceae/química , Bacillus thuringiensis/química , Proteínas de Bactérias/farmacologia , Toxinas Bacterianas/farmacologia , Endotoxinas/farmacologia , Proteínas Hemolisinas/farmacologia , Aedes/crescimento & desenvolvimento , Animais , Toxinas de Bacillus thuringiensis , Sinergismo Farmacológico , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento
9.
Rev Port Cir Cardiotorac Vasc ; 27(3): 231-233, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33068517

RESUMO

Aortoiliac occlusive disease (AIOD) remains an area of debate concerning open and endovascular treatment options. A case of a 63-year old female is reported, with previous known vascular intermittent claudication, that presented in the emergency room with acute ischemia of the right lower limb with 24-hours of evolution. The computer tomographic angiography unveiled occlusion of the superior mesenteric artery, occlusion of left common iliac artery (CIA), subocclusive stenosis of right CIA, occlusion of distal runoffs vessels in the right lower limb and diffuse aorto-iliac disease. The first approach was to place the patient under catheter directed thrombolysis (48h) which led to right pedal pulse recovery but the occlusion of left CIA remained. The patient was then electively submitted to Covered Endovascular Repair of Aortic Bifurcation (CERAB) with chimney to inferior mesenteric artery and with an additional bailout left iliac sandwich due to dissection. Distal pulses are still present after 18 months of follow-up. Endovascular techniques provide a low morbimortality option with similar symptomatic improvement, challenging open surgery as the standard of care even in complex AIOD.


Assuntos
Procedimentos Endovasculares , Artéria Ilíaca/cirurgia , Doença Arterial Periférica , Aorta Abdominal/diagnóstico por imagem , Aorta Abdominal/cirurgia , Angiografia por Tomografia Computadorizada , Feminino , Humanos , Artéria Ilíaca/diagnóstico por imagem , Pessoa de Meia-Idade
10.
Rev Port Cir Cardiotorac Vasc ; 27(1): 23-31, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32239822

RESUMO

Peripheral Arterial Disease (PAD) is a prevalent condition that predisposes the patients to major cardiovascular events. The majority of patients are asymptomatic, however PAD has a great impact in the patients' lifestyle due to its chronic nature. The Peripheral Arterial Questionnaire (PAQ) is a validated tool to quantify the patients' subjective experience of the disease. The aim of this work is to validate the Portuguese version of PAQ. A retrospective study of 59 patients with aortoiliac disease Trans-Atlantic Inter Society Consensus (TASC) type D from two centers in Portugal was conducted. Only 36 patients were able to answer the PAQ and two Portuguese validated questionnaires - a disease-specific (Walk Impairment Questionnaire - WIQ) and a generic one (EuroQol 5 dimensions - 5 level EQ5D-5L). Con- vergent validity of the PAQ was evaluated by correlating the extracted PAQ subscales and Summary score with the WIQ subscales and summary score, as with EQ5D-5L Summary score and EQ5D-5L index by calculating the covariance. The Portuguese version of the peripheral artery questionnaire presented a Cronbach's α for the Summary scale of 0.913. Mean inter-item correlation for the Physical Function domain was 0.471, 0.551 for the Perceived Disability, and 0.464 for Treat- ment Satisfaction. In summary, the Portuguese version of PAQ demonstrated a good level of discrimination between patients with or without symptomatic PAD and its severity and was sensitive to the presence of risk-factors relevant for PAD.


Assuntos
Artérias , Doenças Vasculares Periféricas , Qualidade de Vida , Humanos , Doenças Vasculares Periféricas/complicações , Doenças Vasculares Periféricas/terapia , Portugal , Reprodutibilidade dos Testes , Estudos Retrospectivos , Inquéritos e Questionários
11.
Cell Mol Life Sci ; 75(12): 2211-2226, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29222644

RESUMO

Trichomonas vaginalis is a common sexually transmitted parasite that colonizes the human urogenital tract, where it remains extracellular and adheres to epithelial cells. Infections range from asymptomatic to highly inflammatory, depending on the host and the parasite strain. Despite the serious consequences associated with trichomoniasis disease, little is known about parasite or host factors involved in attachment of the parasite-to-host epithelial cells. Here, we report the identification of microvesicle-like structures (MVs) released by T. vaginalis. MVs are considered universal transport vehicles for intercellular communication as they can incorporate peptides, proteins, lipids, miRNA, and mRNA, all of which can be transferred to target cells through receptor-ligand interactions, fusion with the cell membrane, and delivery of a functional cargo to the cytoplasm of the target cell. In the present study, we demonstrated that T. vaginalis release MVs from the plasma and the flagellar membranes of the parasite. We performed proteomic profiling of these structures demonstrating that they possess physical characteristics similar to mammalian extracellular vesicles and might be selectively charged with specific protein content. In addition, we demonstrated that viable T. vaginalis parasites release large vesicles (LVs), membrane structures larger than 1 µm that are able to interact with other parasites and with the host cell. Finally, we show that both populations of vesicles present on the surface of T vaginalis are induced in the presence of host cells, consistent with a role in modulating cell interactions.


Assuntos
Vesículas Extracelulares/metabolismo , Interações Hospedeiro-Parasita , Vaginite por Trichomonas/metabolismo , Vaginite por Trichomonas/parasitologia , Trichomonas vaginalis/fisiologia , Trichomonas vaginalis/ultraestrutura , Comunicação Celular , Vesículas Extracelulares/química , Vesículas Extracelulares/ultraestrutura , Feminino , Células HeLa , Humanos , Proteômica , Proteínas de Protozoários/análise , Proteínas de Protozoários/metabolismo , Trichomonas vaginalis/química , Trichomonas vaginalis/citologia
12.
J Immunol ; 197(9): 3725-3734, 2016 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-27707998

RESUMO

Acute graft-versus-host disease (aGVHD) is the main complication of allogeneic hematopoietic stem cell transplantation, and many efforts have been made to overcome this important limitation. We showed previously that G-CSF treatment generates low-density splenic granulocytes that inhibit experimental aGVHD. In this article, we show that aGVHD protection relies on incoming IL-10+ neutrophils from G-CSF-treated donor spleen (G-Neutrophils). These G-Neutrophils have high phagocytic capacity, high peroxide production, low myeloperoxidase activity, and low cytoplasmic granule content, which accounts for their low density. Furthermore, they have low expression of MHC class II, costimulatory molecules, and low arginase1 expression. Also, they have low IFN-γ, IL-17F, IL-2, and IL-12 levels, with increased IL-10 production and NO synthase 2 expression. These features are in accordance with the modulatory capacity of G-Neutrophils on regulatory T cell (Treg) generation. In vivo, CD25+ Treg depletion shortly after transplantation with splenic cells from G-CSF-treated donors blocks suppression of aGVHD, suggesting Treg involvement in the protection induced by the G-Neutrophils. The immunocompetence and specificity of the semiallogeneic T cells, long-term after the bone marrow transplant using G-Neutrophils, were confirmed by third-party skin graft rejection; importantly, a graft-versus-leukemia assay showed that T cell activity was maintained, and all of the leukemic cells were eliminated. We conclude that G-CSF treatment generates a population of activated and suppressive G-Neutrophils that reduces aGVHD in an IL-10- and Treg-dependent manner, while maintaining immunocompetence and the graft versus leukemia effect.


Assuntos
Doença Enxerto-Hospedeiro/imunologia , Fator Estimulador de Colônias de Granulócitos/metabolismo , Transplante de Células-Tronco Hematopoéticas , Neutrófilos/imunologia , Linfócitos T Reguladores/imunologia , Doença Aguda , Animais , Células Cultivadas , Doença Enxerto-Hospedeiro/prevenção & controle , Fator Estimulador de Colônias de Granulócitos/imunologia , Tolerância Imunológica , Interleucina-10/metabolismo , Depleção Linfocítica , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Ativação de Neutrófilo , Fagocitose , Espécies Reativas de Oxigênio/metabolismo , Transplante Homólogo
14.
Parasitol Res ; 115(8): 3057-69, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27106236

RESUMO

Proteasomal proteolysis is required for a wide range of cellular processes, including protein quality control, cell cycle progression, cell death and metabolic adaptation to environment changes or stress responses. Proteasome inhibitors are useful compounds for determining the roles of proteasome in eukaryotic cells. Here, we investigated the effects of gliotoxin, a proteasome inhibitor, on the cell growth, replication, ultrastructure, DNA integrity and proteasomal proteolytic activity of the protist parasite Tritrichomonas foetus. The effect of gliotoxin on the transformation of T. foetus to endoflagellar form (EFF), also known as pseudocyst, was investigated. Gliotoxin inhibited the culture growth, arrested cell cycle, and provoked a trichomonacidal effect in a dose-dependent manner. Parasites treated with gliotoxin displayed features typical of cell death, such as membrane blebbing, concentric membrane whorls containing remnants of organelles, intense cytosolic and nuclear vacuolisation, chromatin condensation, DNA fragmentation, cytoplasmic disintegration and plasma membrane disruption. The proteasomal peptidase activity was inhibited by gliotoxin in a dose-dependent manner. Gliotoxin treatment also induced an irreversible EFF transformation in a dose/time-dependent manner. We compared morphological characteristics between gliotoxin- and cold-induced EFF parasites. Our results suggest that gliotoxin could induce EFF transformation by a mechanism distinct from that provoked by cold temperature. This study further contributes to a better understanding of the role of proteasome system in cell cycle, cell death and EFF transformation in T. foetus.


Assuntos
Gliotoxina/farmacologia , Complexo de Endopeptidases do Proteassoma/metabolismo , Inibidores de Proteassoma/farmacologia , Proteólise/efeitos dos fármacos , Tritrichomonas foetus/metabolismo , Animais , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Fragmentação do DNA/efeitos dos fármacos , Tritrichomonas foetus/efeitos dos fármacos
17.
Exp Parasitol ; 142: 67-82, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24793018

RESUMO

The protist parasite Tritrichomonas foetus displays a pear-shaped (PS) and a pseudocystic or endoflagellar form (EFF). Here, we characterised the ecto-phosphatase activity on the surface of EFF and compare its biochemical properties to that of the PS regarding rate of substrate hydrolysis, pH activation profile and sensitivity to well-known phosphatases inhibitors. Two strains exhibiting low- and high-cytotoxicity were used. The enzyme activities of PS and EFF exhibited similar characteristics of protein tyrosine phosphatases (PTP). However, the ecto-phosphatase activities for both forms presented distinct kinetic parameters and different inhibition patterns by PTP inhibitors, suggesting the presence of distinct ecto-enzyme activities between PS and EFF, as well, between both strains. Ultrastructural cytochemistry confirmed the differential distribution of the ecto-phosphatase activity during the EFF transformation. An increase in the percentage of the EFF resulted in a proportional increase in the ecto-phosphatase activity. During EFF reversion, ecto-phosphatase activity decreased and was restored to the level found in the parasites before EFF induction. PS and EFF from the high-cytotoxic strain exhibited higher ecto-phosphatase activities than PS and EFF from the low-cytotoxic strain, respectively. In both strains, the EFF was more cytotoxic and exhibited higher ecto-phosphatase activity when compared to the PS. A large part of the ecto-phosphatase activities of EFF from both strains and PS from the high-cytotoxic strain was irreversibly inhibited when the parasites were pre-treated with a specific antibody against amoebic PTP (anti-EhPRL). Immunoreaction assays revealed that the anti-EhPRL antibody cross-reacted with a 24-kDa protein differentially expressed on the cell surface of PS and EFF T. foetus. A positive correlation was observed between the surface expression of 24-kDa protein and ecto-phosphatase activity. Irreversible inhibition of a part of the ecto-phosphatase activities partially blocked the EFF induction and the cytotoxic effects exerted by both forms. These results suggest that the ecto-phosphatase activities could play a role on the EFF transformation and cytotoxicity of T. foetus.


Assuntos
Fosfoproteínas Fosfatases/metabolismo , Tritrichomonas foetus/enzimologia , Animais , Bovinos , Cães , Imunofluorescência , Histocitoquímica , Concentração de Íons de Hidrogênio , Immunoblotting , Imuno-Histoquímica , Células Madin Darby de Rim Canino , Masculino , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência , Microscopia Imunoeletrônica , Fosfoproteínas Fosfatases/antagonistas & inibidores , Tritrichomonas foetus/classificação , Tritrichomonas foetus/ultraestrutura
18.
Neurocirugia (Astur : Engl Ed) ; 35(2): 71-78, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-37696419

RESUMO

INTRODUCTION AND OBJECTIVES: Carotid cross-clamping during carotid endarterectomy might lead to intraoperative neurologic deficits, increasing stroke/death risk. If deficits are detected, carotid shunting has been recommended to reduce the risk of stroke. However, shunting may sustain a specific chance of embolic events and subsequently incurring harm. Current evidence is still questionable regarding its clear benefit. The aim is to determine whether a policy of selective shunt impacts the complication rate following an endarterectomy. MATERIAL AND METHODS: From January 2013 to May 2021, all patients undergoing carotid endarterectomy under regional anesthesia with intraoperative neurologic alteration were retrieved. Patients submitted to selective shunt were compared to a non-shunt group. A 1:1 propensity score matching (PSM) was performed. Differences between the groups and clinical outcomes were calculated, resorting to univariate analysis. RESULTS: Ninety-eight patients were selected, from which 23 were operated on using a shunt. After PSM, 22 non-shunt patients were compared to 22 matched shunted patients. Concerning demographics and comorbidities, both groups were comparable to pre and post-PSM, except for chronic heart failure, which was more prevalent in shunted patients (26.1%, P=0.036) in pre-PSM analysis. Regarding 30-day stroke and score Clavien-Dindo ≥2, no significant association was found (P=0.730, P=0.635 and P=0.942, P=0.472, correspondingly, for pre and post-PSM). CONCLUSIONS: In this cohort, resorting to shunting did not demonstrate an advantage regarding 30-day stroke or a Clavien-Dindo ≥ 2 rates. Nevertheless, additional more extensive studies are mandatory to achieve precise results concerning the accurate utility of carotid shunting in this subset of patients under regional anesthesia.


Assuntos
Estenose das Carótidas , Endarterectomia das Carótidas , Acidente Vascular Cerebral , Humanos , Endarterectomia das Carótidas/efeitos adversos , Endarterectomia das Carótidas/métodos , Estenose das Carótidas/cirurgia , Estenose das Carótidas/complicações , Pontuação de Propensão , Fatores de Risco , Acidente Vascular Cerebral/epidemiologia , Acidente Vascular Cerebral/etiologia
19.
Elife ; 122023 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-37129369

RESUMO

Trichomonas vaginalis, the etiologic agent of the most common non-viral sexually transmitted infection worldwide. With an estimated annual prevalence of 276 million new cases, mixed infections with different parasite strains are expected. Although it is known that parasites interact with their host to enhance their own survival and transmission, evidence of mixed infections call into question the extent to which unicellular parasites communicate with each other. Here, we demonstrated that different T. vaginalis strains can communicate through the formation of cytoneme-like membranous cell connections. We showed that cytonemes formation of an adherent parasite strain (CDC1132) is affected in the presence of a different strain (G3 or B7RC2). Our findings provide evidence that this effect is contact-independent and that extracellular vesicles (EVs) are responsible, at least in part, of the communication among strains. We found that EVs isolated from G3, B7RC2, and CDC1132 strains contain a highly distinct repertoire of proteins, some of them involved in signaling and communication, among other functions. Finally, we showed that parasite adherence to host cells is affected by communication between strains as binding of adherent T. vaginalis CDC1132 strain to prostate cells is significantly higher in the presence of G3 or B7RC2 strains. We also observed that a poorly adherent parasite strain (G3) adheres more strongly to prostate cells in the presence of an adherent strain. The study of signaling, sensing, and cell communication in parasitic organisms will enhance our understanding of the basic biological characteristics of parasites, which may have important consequences in pathogenesis.


Assuntos
Coinfecção , Vesículas Extracelulares , Parasitos , Trichomonas vaginalis , Masculino , Animais , Humanos , Trichomonas vaginalis/metabolismo , Vesículas Extracelulares/metabolismo , Comunicação Celular
20.
J Cardiovasc Surg (Torino) ; 64(1): 48-57, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36168948

RESUMO

BACKGROUND: Aortoiliac peripheral artery disease may lead to disabling lower limb claudication or to lower limb chronic threatening ischemia, which is associated with increased short and long-term morbi-mortality. The red blood cell distribution width-coefficient of variation (RDW-CV) has been able to predict outcomes in other atherosclerotic diseases, such as myocardial infarction and stroke. The main objective of this study was to assess the predictive ability of perioperative RDW-CV in accurately predicting short and long-term major adverse cardiovascular events (MACE) and all-cause mortality in patients submitted to aortoiliac revascularization due to extensive aortoiliac atherosclerotic disease. METHODS: From 2013 to 2020, patients who underwent aortoiliac revascularization due to severe aortoiliac disease were included in a prospective cohort. Blood samples were taken preoperatively and the patient's demographics, comorbidities, and postoperative outcomes were assessed. A multivariate Cox regression model was used to adjust for confounding and assess the independent effect of these prognostic factors on the outcomes. RESULTS: The study group included 107 patients. Median follow-up was 57 (95% CI: 34.4-69.6) months. Preoperative RDW-CV was increased in thirty-eight patients (35.5%). Increased RDW-CV was associated with congestive heart failure - adjusted odds ratio of 5.043 (95% CI: 1.436-17.717, P=0.012). It could predict long-term occurrence of MACE (adjusted hazard ratio [aHR] 1.065, 95% CI: 1.014-1.118, P=0.011), all-cause mortality (aHR=1.069, 95% CI: 1.014-1.126, P=0.013), acute heart failure (AHF) (aHR=1.569, 95% CI: 1.179-2.088, P=0.002), and stroke (aHR=1.343, 95% CI: 1.044-1.727, P=0.022). CONCLUSIONS: RDW is a widely available and low-cost marker that was able to independently predict long-term AHF, stroke, MACE, and all-cause mortality in patients with extensive aortoiliac disease submitted to revascularization. This biomarker could help assess which patients would likely benefit from stricter follow-up in the long-term.


Assuntos
Insuficiência Cardíaca , Acidente Vascular Cerebral , Humanos , Prognóstico , Estudos Prospectivos , Índices de Eritrócitos , Acidente Vascular Cerebral/epidemiologia , Eritrócitos , Fatores de Risco
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