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Extracellular vesicles (EVs) play a crucial role in intercellular communication by transferring bioactive molecules from donor to recipient cells. As a result, EV fusion leads to the modulation of cellular functions and has an impact on both physiological and pathological processes in the recipient cell. This study explores the impact of EV fusion on cellular responses to inflammatory signaling. Our findings reveal that fusion renders non-responsive cells susceptible to inflammatory signaling, as evidenced by increased NF-κB activation and the release of inflammatory mediators. Syntaxin-binding protein 1 is essential for the merge and activation of intracellular signaling. Subsequent analysis show that EVs transfer their functionally active receptors to target cells, making them prone to an otherwise unresponsive state. EVs in complex with their agonist, require no further stimulation of the target cells to trigger mobilization of NF-κB. While receptor antagonists were unable to inhibit NF-κB activation, blocking of the fusion between EVs and their target cells with heparin mitigated inflammation in mice challenged with EVs.
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Vesículas Extracelulares , NF-kappa B , Animais , Camundongos , NF-kappa B/metabolismo , Vesículas Extracelulares/metabolismo , Transporte Biológico , Transdução de Sinais , Inflamação/patologiaRESUMO
PURPOSE: A pilot study to identify bacterial biofilm in the lacrimal sacs of patients with chronic dacryocystitis, and in patients with epiphora but without discharge, using scanning electron microscopy. METHODS: Five patients: two with nasolacrimal duct obstruction without dacryocystitis, and three with dacryocystitis refractory to antibiotics, underwent external dacryocystorhinostomy. One control patient without infection was included. Bacterial cultures were obtained from the lumen of the lacrimal sac to analyze possible bacterial growth, including antibiotic resistance. Biopsies were taken from all lacrimal sacs and prepared for light and scanning electron microscopy. RESULTS: Scanning electron microscopy of all the lacrimal sac samples revealed structures consistent with bacterial communities and adjacent extracellular material, indicating biofilm formation. This was most prominent in one of the patients with chronic dacryocystitis. Bacteria were found not only on the luminal surface of the sac, but also within the tissue of the sac. Bacterial growth was identified in samples from two patients with chronic dacryocystitis, whereas samples from the other three patients showed no bacterial growth. CONCLUSION: Lack of patency of the lacrimal duct predisposes to bacterial growth, even in patients with no clinically confirmed infection of the lacrimal sac. The finding of a biofilm in patients with chronic dacryocystitis explains the lack of efficiency of antibiotic treatment at the concentrations used in clinical practice.
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Dacriocistite , Dacriocistorinostomia , Obstrução dos Ductos Lacrimais , Ducto Nasolacrimal , Humanos , Obstrução dos Ductos Lacrimais/terapia , Obstrução dos Ductos Lacrimais/complicações , Projetos Piloto , Dacriocistite/cirurgia , Dacriocistorinostomia/efeitos adversos , Bactérias , Antibacterianos/uso terapêuticoRESUMO
[This corrects the article DOI: 10.3389/fimmu.2018.01988.].
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Achromobacter is a genus of Gram-negative rods, which can cause persistent airway infections in people with cystic fibrosis (CF). The knowledge about virulence and clinical implications of Achromobacter is still limited, and it is not fully established whether Achromobacter infections contribute to disease progression or if it is a marker of poor lung function. The most commonly reported Achromobacter species in CF is A. xylosoxidans. While other Achromobacter spp. are also identified in CF airways, the currently used Matrix-Assisted Laser Desorption/Ionization Time Of Flight Mass Spectrometry (MALDI-TOF MS) method in routine diagnostics cannot distinguish between species. Differences in virulence between Achromobacter species have consequently not been well studied. In this study, we compare phenotypes and proinflammatory properties of A. xylosoxidans, A. dolens, A. insuavis, and A. ruhlandii using in vitro models. Bacterial supernatants were used to stimulate CF bronchial epithelial cells and whole blood from healthy individuals. Supernatants from the well-characterized CF-pathogen Pseudomonas aeruginosa were included for comparison. Inflammatory mediators were analyzed with ELISA and leukocyte activation was assessed using flow cytometry. The four Achromobacter species differed in morphology seen in scanning electron microscopy (SEM), but there were no observed differences in swimming motility or biofilm formation. Exoproducts from all Achromobacter species except A. insuavis caused significant IL-6 and IL-8 secretion from CF lung epithelium. The cytokine release was equivalent or stronger than the response induced by P. aeruginosa. All Achromobacter species activated neutrophils and monocytes ex vivo in a lipopolysaccharide (LPS)-independent manner. Our results indicate that exoproducts of the four included Achromobacter species do not differ consistently in causing inflammatory responses, but they are equally or even more capable of inducing inflammation compared with the classical CF pathogen P. aeruginosa. IMPORTANCE Achromobacter xylosoxidans is an emerging pathogen among people with cystic fibrosis (CF). Current routine diagnostic methods are often unable to distinguish A. xylosoxidans from other Achromobacter species, and the clinical relevance of different species is still unknown. In this work, we show that four different Achromobacter species relevant to CF evoke similar inflammatory responses from airway epithelium and leukocytes in vitro, but they are all equally or even more proinflammatory compared to the classic CF-pathogen Pseudomonas aeruginosa. The results suggest that Achromobacter species are important airway pathogens in CF, and that all Achromobacter species are relevant to treat.
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Achromobacter denitrificans , Achromobacter , Fibrose Cística , Infecções por Bactérias Gram-Negativas , Humanos , Achromobacter/genética , Fibrose Cística/complicações , Fibrose Cística/microbiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Achromobacter denitrificans/genética , PulmãoRESUMO
Biofilm formation by Streptococcus pyogenes (group A streptococcus [GAS]) in model systems mimicking the respiratory tract is poorly documented. Most studies have been conducted on abiotic surfaces, which poorly represent human tissues. We have previously shown that GAS forms mature and antibiotic-resistant biofilms on physiologically relevant epithelial cells. However, the roles of the substratum, extracellular matrix (ECM) components, and GAS virulence factors in biofilm formation and structure are unclear. In this study, biofilm formation was measured on respiratory epithelial cells and keratinocytes by determining biomass and antibiotic resistance, and biofilm morphology was visualized using scanning electron microscopy. All GAS isolates tested formed biofilms that had similar, albeit not identical, biomass and antibiotic resistance for both cell types. Interestingly, functionally mature biofilms formed more rapidly on keratinocytes but were structurally denser and coated with more ECM on respiratory epithelial cells. The ECM was crucial for biofilm integrity, as protein- and DNA-degrading enzymes induced bacterial release from biofilms. Abiotic surfaces supported biofilm formation, but these biofilms were structurally less dense and organized. No major role for M protein, capsule, or streptolysin O was observed in biofilm formation on epithelial cells, although some morphological differences were detected. NAD-glycohydrolase was required for optimal biofilm formation, whereas streptolysin S and cysteine protease SpeB impaired this process. Finally, no correlation was found between cell adherence or autoaggregation and GAS biofilm formation. Combined, these results provide a better understanding of the role of biofilm formation in GAS pathogenesis and can potentially provide novel targets for future treatments against GAS infections.
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Biofilmes/crescimento & desenvolvimento , Células Epiteliais/microbiologia , Streptococcus pyogenes/patogenicidade , Fatores de Virulência/metabolismo , Antibacterianos/farmacologia , Antígenos de Bactérias/genética , Antígenos de Bactérias/metabolismo , Cápsulas Bacterianas/genética , Cápsulas Bacterianas/metabolismo , Proteínas da Membrana Bacteriana Externa/genética , Proteínas da Membrana Bacteriana Externa/metabolismo , Biofilmes/efeitos dos fármacos , Biomassa , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Farmacorresistência Bacteriana , Células Epiteliais/ultraestrutura , Matriz Extracelular/microbiologia , Matriz Extracelular/ultraestrutura , Queratinócitos/microbiologia , Queratinócitos/ultraestrutura , Sorogrupo , Fatores de Virulência/genéticaRESUMO
[This corrects the article DOI: 10.3389/fphys.2016.00330.].
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Introduction: As medical advances are achieved in the care of chronically ill patients, there is increasing evidence that health-related quality of life (QoL) is associated with poor outcomes, including hospitalization and death. This study aimed to evaluate QoL as a predictor of hospitalization and death in patients with chronic kidney disease (CKD) on hemodialysis. Methods: A retrospective cohort study of 108 patients with CKD on hemodialysis with 24-month follow-up. QoL was assessed by the Kidney Disease Quality of Life Short-Form (KDQOL-SF), including time to first hospitalization and death as outcomes. Results: The highest KDQOL-SF scores at baseline were observed in Sexual function, Dialysis staff encouragement, and Cognitive function, while the lowest scores were observed in Working status, Role physical, and Energy/fatigue. There was an association of Overall health and Role emotional domains with shorter time to first hospitalization. Data analyzed were insufficient to indicate an association of QoL with mortality in this population. Conclusion: QoL was associated with time to first hospitalization in patients with CKD on hemodialysis, but the results were not sufficient to indicate its association with mortality. (AU)
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Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Qualidade de Vida , Insuficiência Renal Crônica/complicações , Insuficiência Renal Crônica/epidemiologia , Diálise Renal/efeitos adversosRESUMO
Non-typeable Haemophilus influenzae (NTHi) is a Gram-negative human commensal commonly residing in the nasopharynx of preschool children. It occasionally causes upper respiratory tract infection such as acute otitis media, but can also spread to the lower respiratory tract causing bronchitis and pneumonia. There is increasing recognition that NTHi has an important role in chronic lower respiratory tract inflammation, particularly in persistent infection in patients suffering from chronic obstructive pulmonary disease (COPD). Here, we set out to assess the innate protective effects of collagen VI, a ubiquitous extracellular matrix component, against NTHi infection in vivo. In vitro, collagen VI rapidly kills bacteria through pore formation and membrane rupture, followed by exudation of intracellular content. This effect is mediated by specific binding of the von Willebrand A (VWA) domains of collagen VI to the NTHi surface adhesins protein E (PE) and Haemophilus autotransporter protein (Hap). Similar observations were made in vivo specimens from murine airways and COPD patient biopsies. NTHi bacteria adhered to collagen fibrils in the airway mucosa and were rapidly killed by membrane destabilization. The significance in host-pathogen interplay of one of these molecules, PE, was highlighted by the observation that it confers partial protection from bacterial killing. Bacteria lacking PE were more prone to antimicrobial activity than NTHi expressing PE. Altogether the data shed new light on the carefully orchestrated molecular events of the host-pathogen interplay in COPD and emphasize the importance of the extracellular matrix as a novel branch of innate host defense.
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Colágeno Tipo IV/metabolismo , Matriz Extracelular/metabolismo , Fibroblastos/fisiologia , Infecções por Haemophilus/imunologia , Haemophilus influenzae/fisiologia , Pulmão/imunologia , Doença Pulmonar Obstrutiva Crônica/imunologia , Adesinas Bacterianas/metabolismo , Animais , Células Cultivadas , Modelos Animais de Doenças , Feminino , Infecções por Haemophilus/complicações , Interações Hospedeiro-Patógeno , Humanos , Evasão da Resposta Imune , Imunidade Inata , Camundongos , Camundongos Endogâmicos C57BL , Ligação Proteica , Doença Pulmonar Obstrutiva Crônica/complicaçõesRESUMO
Collagen VI is a ubiquitous extracellular matrix component that forms extensive microfibrillar networks in most connective tissues. In this study, we describe for the first time, to our knowledge, that the collagen VI von Willebrand factor type A-like domains exhibit a broad-spectrum antimicrobial activity against Gram-positive and Gram-negative bacteria in human skin infections in vivo. In silico sequence and structural analysis of VWA domains revealed that they contain cationic and amphipathic peptide sequence motifs, which might explain the antimicrobial nature of collagen VI. In vitro and in vivo studies show that these peptides exhibited significant antibacterial activity against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa through membrane disruption. Our findings shed new light on the role of collagen VI-derived peptides in innate host defense and provide templates for development of peptide-based antibacterial therapies.
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Antibacterianos/imunologia , Colágeno Tipo VI/imunologia , Peptídeos/imunologia , Bactérias/imunologia , Infecções Bacterianas/imunologia , Humanos , Imunidade Inata/imunologia , Domínios Proteicos/imunologia , Pele/imunologia , Pele/microbiologia , Dermatopatias Bacterianas/imunologiaRESUMO
Severe cerebral intraventricular hemorrhage (IVH) in preterm infants continues to be a major clinical problem, occurring in about 15-20% of very preterm infants. In contrast to other brain lesions the incidence of IVH has not been reduced over the last decade, but actually slightly increased. Currently over 50% of surviving infants develop post-hemorrhagic ventricular dilatation and about 35% develop severe neurological impairment, mainly cerebral palsy and intellectual disability. To date there is no therapy available to prevent infants from developing either hydrocephalus or serious neurological disability. It is known that blood rapidly accumulates within the ventricles following IVH and this leads to disruption of normal anatomy and increased local pressure. However, the molecular mechanisms causing brain injury following IVH are incompletely understood. We propose that extracellular hemoglobin is central in the pathophysiology of periventricular white matter damage following IVH. Using a preterm rabbit pup model of IVH the distribution of extracellular hemoglobin was characterized at 72 h following hemorrhage. Evaluation of histology, histochemistry, hemoglobin immunolabeling and scanning electron microscopy revealed presence of extensive amounts of extracellular hemoglobin, i.e., not retained within erythrocytes, in the periventricular white matter, widely distributed throughout the brain. Furthermore, double immunolabeling together with the migration and differentiation markers polysialic acid neural cell adhesion molecule (PSA-NCAM) demonstrates that a significant proportion of the extracellular hemoglobin is distributed in areas of the periventricular white matter with high extracellular plasticity. In conclusion, these findings support that extracellular hemoglobin may contribute to the pathophysiological processes that cause irreversible damage to the immature brain following IVH.
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Moraxella catarrhalis is a Gram-negative human mucosal commensal and pathogen. It is a common cause of exacerbation in chronic obstructive pulmonary disease (COPD). During the process of infection, host colonization correlates with recognition of host molecular patterns. Importantly, in COPD patients with compromised epithelial integrity the underlying extracellular matrix is exposed and provides potential adhesive targets. Collagen VI is a ubiquitous fibrillar component in the airway mucosa and has been attributed both adhesive and killing properties against Gram-positive bacteria. However, less is known regarding Gram-negative microorganisms. Therefore, in the present study, the interaction of M. catarrhalis with collagen VI was characterized. We found that collagen VI is upregulated in the airways of COPD patients and exposed upon epithelial desquamation. Ex vivo, we inoculated airway biopsies and fibroblasts from COPD patients with M. catarrhalis. The bacteria specifically adhered to collagen VI-containing matrix fibrils. In vitro, purified collagen VI microfibrils bound to bacterial surface structures. The primary adhesion target was mapped to the collagen VI α2-chain. Upon exposure to collagen VI, bacteria were killed by membrane destabilization in physiological conditions. These previously unknown properties of collagen VI provide novel insights into the extracellular matrix innate immunity by quickly entrapping and killing pathogen intruders.
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Colágeno Tipo VI/metabolismo , Moraxella catarrhalis/imunologia , Infecções por Moraxellaceae/imunologia , Mucosa Nasal/imunologia , Doença Pulmonar Obstrutiva Crônica/imunologia , Aderência Bacteriana , Bacteriólise , Células Cultivadas , Colágeno Tipo VI/genética , Matriz Extracelular/metabolismo , Fibroblastos/imunologia , Fibroblastos/microbiologia , Interações Hospedeiro-Patógeno , Humanos , Imunidade Inata , Infecções por Moraxellaceae/complicações , Mucosa Nasal/microbiologia , Doença Pulmonar Obstrutiva Crônica/complicações , Regulação para CimaRESUMO
Platelet activation and aggregation have been reported to occur in response to a number of Gram-positive pathogens. Here, we show that platelet aggregates induced by Streptococcus pyogenes were unstable and that viable bacteria escaped from the aggregates over time. This was not due to differential activation in response to the bacteria compared with physiological activators. All the bacterial isolates induced significant platelet activation, including integrin activation and alpha and dense-granule release, at levels equivalent to those induced by potent physiological platelet activators that induced stable aggregates. The ability to escape the aggregates and to resist the antibacterial effects of platelets was dependent on active protein synthesis by the bacteria within the aggregate. We conclude that S. pyogenes bacteria can temporarily cover themselves with activated platelets, and we propose that this may facilitate survival of the bacteria in the presence of platelets.
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Aderência Bacteriana , Plaquetas/microbiologia , Interações Hospedeiro-Patógeno , Ativação Plaquetária , Agregação Plaquetária , Streptococcus pyogenes/fisiologia , Adulto , Feminino , Humanos , MasculinoRESUMO
The innate immune factors controlling Candida albicans are mostly unknown. Vulvovaginal candidiasis is common in women and affects approximately 70-75% of all women at least once. Despite the propensity of Candida to colonize the vagina, transmission of Candida albicans following sexual intercourse is very rare. This prompted us to investigate whether the post coital vaginal milieu contained factors active against C. albicans. By CFU assays, we found prominent candidacidal activity of post coital seminal plasma at both neutral and the acid vaginal pH. In contrast, normal seminal plasma did not display candidacidal activity prior to acidification. By antifungal gel overlay assay, one clearing zone corresponding to a protein band was found in both post coital and normal seminal plasma, which was subsequently identified as ß-microseminoprotein. At neutral pH, the fungicidal activity of ß-microseminoprotein and seminal plasma was inhibited by calcium. By NMR spectroscopy, amino acid residue E(71) was shown to be critical for the calcium coordination. The acidic vaginal milieu unleashed the fungicidal activity by decreasing the inhibitory effect of calcium. The candidacidal activity of ß-microseminoprotein was mapped to a fragment of the C-terminal domain with no structural similarity to other known proteins. A homologous fragment from porcine ß-microseminoprotein demonstrated calcium-dependent fungicidal activity in a CFU assay, suggesting this may be a common feature for members of the ß-microseminoprotein family. By electron microscopy, ß-microseminoprotein was found to cause lysis of Candida. Liposome experiments demonstrated that ß-microseminoprotein was active towards ergosterol-containing liposomes that mimic fungal membranes, offering an explanation for the selectivity against fungi. These data identify ß-microseminoprotein as an important innate immune factor active against C. albicans and may help explain the low sexual transmission rate of Candida.
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Antifúngicos/imunologia , Cálcio/imunologia , Candida albicans/imunologia , Coito , Imunidade Inata , Proteínas Secretadas pela Próstata/imunologia , Sêmen/imunologia , Antifúngicos/química , Cálcio/química , Candida albicans/patogenicidade , Candidíase/imunologia , Feminino , Humanos , Concentração de Íons de Hidrogênio , Lipossomos/química , Lipossomos/imunologia , Masculino , Proteínas Secretadas pela Próstata/química , Estrutura Terciária de Proteína , Sêmen/química , Vagina/imunologia , Vagina/metabolismoRESUMO
Collagen type VI is a subepithelial extracellular matrix component in airways and an adhesive substrate for oral pathogens [Bober et al.: J Innate Immun 2010;2:160-166]. Here, we report that collagen VI displays a dose-dependent antimicrobial activity against group A, C, and G streptococci by membrane disruption in physiological conditions. The data disclose previously unrecognized aspects of the extracellular matrix in innate host defense.
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Anti-Infecciosos/farmacologia , Colágeno Tipo VI/farmacologia , Matriz Extracelular/imunologia , Streptococcus pyogenes/efeitos dos fármacos , Streptococcus/efeitos dos fármacos , Anti-Infecciosos/química , Anti-Infecciosos/metabolismo , Colágeno Tipo VI/química , Colágeno Tipo VI/metabolismo , Matriz Extracelular/química , Humanos , Imunidade Inata , Testes de Sensibilidade Microbiana , Streptococcus/classificaçãoRESUMO
As infecções no trato urinário (ITUs) representam a forma mais comum de infecções bacterianas em gestantes. A gravidez é uma situação que predispõe ao aparecimento das ITUs, devido às mudanças fisiológicas (mecânicas e hormonais) que ocorrem nesse período da vida da mulher, facilitando à transformação das mulheres bacteriúricas assintomáticas (BA) em sintomáticas. A BA acomete entre 2-10% das gestantes, se não tratadas adequadamente podem desenvolver pielonefrite em 40% dos casos. As ITUs manifestam-se clinicamente por disúria, polaciúria, urgência miccional e dor no baixo ventre na cistite, arrepios de frio e lombalgia na pielonefrite, ou completa ausência de sintomas na bacteriúria assintomática. O diagnóstico, na maioria das vezes, com exceção da bacteriúria assintomática, é clínico. Deve-se levar em consideração fatores, como a condição da paciente, a tolerabilidade e a toxicidade materna e fetal para a escolha da melhor abordagem terapêutica. O objetivo do presente estudo foi revisar os trabalhos que abordassem fisiopatologia, formas clínicas das ITUs, epidemiologia, etiologia, métodos de diagnóstico e tratamento das infecções urinárias na gestação.
The urinary tract infections (UTIs) represent the most common form of bacterial infections in pregnant women. Pregnancy is a situation that predisposes to the development of UTIs due to physiological changes (mechanical and hormonal) that occur in this period of women?s life, facilitating the transformation of asymptomatic bacteriuria women (AB) in symptomatic ones. AB affects 2-10% of pregnant women, and if not properly treated, it can develop pyelonephritis in 40% of cases. UTIs are clinically manifested by dysuria, urinary frequency, urinary urgency and lower abdominal pain in cystitis, chills and low back pain in pyelonephritis, or complete absence of symptoms in asymptomatic bacteriuria. The diagnosis in most cases is clinical, except for asymptomatic bacteriuria. One should take into account factors such as the patient?s condition, tolerability, and maternal and fetal toxicity when choosing the best therapeutic approach. The aim of this study was to review the work that addressed pathophysiology, clinical forms of UTIs, epidemiology, etiology, diagnostic methods and treatment of urinary infections in pregnancy.
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Epithelial linings serve as physical barriers and produce antimicrobial peptides (AMPs) to maintain host integrity. Examples are the bactericidal proteins midkine (MK) and BRAK/CXCL14 that are constitutively produced in the skin epidermal layer, where the anaerobic Gram-positive coccoid commensal Finegoldia magna resides. Consequently, this bacterium is likely to encounter both MK and BRAK/CXCL14, making these molecules possible threats to its habitat. In this study, we show that MK expression is upregulated during inflammation, concomitant with a strong downregulation of BRAK/CXCL14, resulting in changed antibacterial conditions. MK, BRAK/CXCL14, and the inflammation-dependent antimicrobial ß-defensins human ß-defensin (hBD)-2 and hBD-3 all showed bactericidal activity against both F. magna and the virulent pathogen Streptococcus pyogenes at similar concentrations. SufA, a released protease of F. magna, degraded MK and BRAK/CXCL14 but not hBD-2 nor hBD-3. Cleavage was seen at lysine and arginine residues, amino acids characteristic of AMPs. Intermediate SufA-degraded fragments of MK and BRAK/CXCL14 showed stronger bactericidal activity against S. pyogenes than F. magna, thus promoting survival of the latter. In contrast, the cysteine-protease SpeB of S. pyogenes rapidly degraded all AMPs investigated. The proteins FAF and SIC, released by F. magna and S. pyogenes, respectively, neutralized the antibacterial activity of MK and BRAK/CXCL14, protein FAF being the most efficient. Quantitation and colocalization by immunoelectron microscopy demonstrated significant levels and interactions of the molecules in in vivo and ex vivo samples. The findings reflect strategies used by a permanently residing commensal and a virulent pathogen, the latter operating during the limited time course of invasive disease.
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Peptídeos Catiônicos Antimicrobianos/metabolismo , Epitélio/imunologia , Epitélio/microbiologia , Inflamação/metabolismo , Streptococcus pyogenes/patogenicidade , Células Cultivadas , Eletroforese em Gel de Poliacrilamida , Epitélio/metabolismo , Bactérias Gram-Positivas/metabolismo , Humanos , Imuno-Histoquímica , Inflamação/imunologia , Espectrometria de Massas , Microscopia Eletrônica de Transmissão , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Streptococcus pyogenes/metabolismo , Ressonância de Plasmônio de SuperfícieRESUMO
Objetivo: Avaliar o comportamento da função pulmonar e da dor em pacientes adultos submetidos à cirurgia cardíaca por esternotomia. Além de verificar possíveis correlações e comparações dessas variáveis com as características do procedimento cirúrgico e o tempo de internação hospitalar. Métodos: Foi realizado estudo de coorte composto de 70 indivíduos, nos quais foi avaliada a função pulmonar préoperatória por espirometria e inspirometria de incentivo. Os pacientes foram acompanhados no pós-operatório, por meio de protocolo com informações da cirurgia, função pulmonar e um protocolo de avaliação álgica (escala análoga visual e desenho do corpo humano). Resultados: Os valores de função pulmonar do período pós-operatório apresentaram diminuição significativa em relação ao pré-operatório (P<0,01). A dor localizou-se na região da esternotomia, persistindo até o 5º dia de pósoperatório. Houve correlação da dor com os parâmetros de função pulmonar (volume expiratório forçado no 1º segundo - percentual r=-0,271 e P<0,047; pico de fluxo expiratório r=-0,357 e P<0,008; volume inspiratório máximo r=-0,293 e P<0,032). Não se observou correlação significativa da dor com outras variáveis. Conclusão: Observou-se prejuízo significativo da função pulmonar, não se restabelecendo completamente até o 5º dia de pós-operatório. A dor foi uma queixa que persistiu durante todo o período do estudo. Os parâmetros de função pulmonar apresentaram relação significativa com a dor. Não houve correlação entre dor e as características dos indivíduos, do procedimento cirúrgico e tempo de internação hospitalar.
Objective: To investigate the pulmonary function and pain in adult patients undergoing heart surgery via sternotomy and to verify possible correlations of these variables with the characteristics of the surgical procedure and hospital stay. Methods: A cross-sectional study was carried out of 70 individuals undergoing heart surgery. The lung function was assessed before and after surgery by spirometry and incentive spirometry. Details of the surgical procedure were studied and patients were followed up postoperatively using a visual analogue scale and design of the human body to evaluate pain. Results: The pulmonary function was significantly impaired in the postoperative compared to preoperative period (P <0.01). The pain was centered in the region of the sternotomy and persisted until at least the 5th postoperative day. There was a correlation between pain and the parameters of pulmonary function (forced expiratory volume in 1 second - percentage: r = -0.271, P <0.047; peak expiratory flow: r = 0.357, P <0.008; and maximum inspiratory volume: r = -0.293, P <0.032). There was no significant correlation between pain and other variables. Conclusion: There was significant impairment of lung function which had not recovered completely on the 5th postoperative day. Pain was a complaint that persisted throughout the study period. The parameters of pulmonary function showed a significant relationship with pain. There was no correlation between pain and the characteristics of individuals, the surgical procedure or the length of hospital stay.
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Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Procedimentos Cirúrgicos Cardíacos/métodos , Pulmão/fisiopatologia , Dor Pós-Operatória/fisiopatologia , Esternotomia/efeitos adversos , Métodos Epidemiológicos , Tempo de Internação , Medição da Dor/métodos , Fatores de TempoRESUMO
OBJECTIVE: To investigate the pulmonary function and pain in adult patients undergoing heart surgery via sternotomy and to verify possible correlations of these variables with the characteristics of the surgical procedure and hospital stay. METHODS: A cross-sectional study was carried out of 70 individuals undergoing heart surgery. The lung function was assessed before and after surgery by spirometry and incentive spirometry. Details of the surgical procedure were studied and patients were followed up postoperatively using a visual analogue scale and design of the human body to evaluate pain. RESULTS: The pulmonary function was significantly impaired in the postoperative compared to preoperative period (P <0.01). The pain was centered in the region of the sternotomy and persisted until at least the 5th postoperative day. There was a correlation between pain and the parameters of pulmonary function (forced expiratory volume in 1 second - percentage: r = -0.271, P <0.047; peak expiratory flow: r = 0.357, P <0.008; and maximum inspiratory volume: r = -0.293, P <0.032). There was no significant correlation between pain and other variables. CONCLUSION: There was significant impairment of lung function which had not recovered completely on the 5th postoperative day. Pain was a complaint that persisted throughout the study period. The parameters of pulmonary function showed a significant relationship with pain. There was no correlation between pain and the characteristics of individuals, the surgical procedure or the length of hospital stay.
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Procedimentos Cirúrgicos Cardíacos/métodos , Pulmão/fisiopatologia , Dor Pós-Operatória/fisiopatologia , Esternotomia/efeitos adversos , Métodos Epidemiológicos , Feminino , Humanos , Tempo de Internação , Masculino , Pessoa de Meia-Idade , Medição da Dor/métodos , Fatores de TempoRESUMO
Caveolin-1 (Cav-1) is essential for the morphology of membrane caveolae and exerts a negative influence on a number of signaling systems, including nitric oxide (NO) production and activity of the MAP kinase cascade. In the vascular system, ablation of caveolin-1 may thus be expected to cause arterial dilatation and increased vessel wall mass (remodeling). This was tested in Cav-1 knockout (KO) mice by a detailed morphometric and functional analysis of mesenteric resistance arteries, shown to lack caveolae. Quantitative morphometry revealed increased media thickness and media-to-lumen ratio in KO. Pressure-induced myogenic tone and flow-induced dilatation were decreased in KO arteries, but both were increased toward wild-type (WT) levels following NO synthase (NOS) inhibition. Isometric force recordings following NOS inhibition showed rightward shifts of passive and active length-force relationships in KO, and the force response to alpha(1)-adrenergic stimulation was increased. In contrast, media thickness and force response of the aorta were unaltered in KO vs. WT, whereas lumen diameter was increased. Mean arterial blood pressure during isoflurane anesthesia was not different in KO vs. WT, but greater fluctuation in blood pressure over time was noted. Following NOS inhibition, fluctuations disappeared and pressure increased twice as much in KO (38 +/- 6%) compared with WT (17 +/- 3%). Tracer-dilution experiments showed increased plasma volume in KO. We conclude that NO affects blood pressure more in Cav-1 KO than in WT mice and that restructuring of resistance vessels and an increased responsiveness to adrenergic stimulation compensate for a decreased tone in Cav-1 KO mice.
Assuntos
Caveolina 1/deficiência , Artérias Mesentéricas/fisiologia , Substitutos do Plasma/farmacologia , Volume Plasmático/fisiologia , Animais , Pressão Sanguínea/fisiologia , Western Blotting , Caveolina 1/genética , Inibidores Enzimáticos/farmacologia , Imunofluorescência , Contração Isométrica/efeitos dos fármacos , Cinética , Artérias Mesentéricas/citologia , Artérias Mesentéricas/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microscopia Eletrônica de Transmissão , Contração Muscular/efeitos dos fármacos , Contração Muscular/fisiologia , Tono Muscular/fisiologia , Miografia , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico Sintase Tipo III/antagonistas & inibidores , Técnicas de Cultura de Órgãos , Volume Plasmático/efeitos dos fármacos , Timidina/metabolismo , Vasodilatação/fisiologiaRESUMO
The ability of protein kinase C and casein kinase 2 substrate in neurons (PACSIN)/syndapin proteins to self-polymerize is crucial for the simultaneous interactions with more than one Src homology 3 domain-binding partner or with lipid membranes. The assembly of this network has profound effects on the neural Wiskott-Aldrich syndrome protein-mediated attachment of the actin polymerization machinery to vesicle membranes as well as on the movement of the corresponding vesicles. Also, the sensing of vesicle membranes and/or the induction of membrane curvature are more easily facilitated in the presence of larger PACSIN complexes. The N-terminal Fes-CIP homology and Bin-Amphiphysin-Rvs (F-BAR) domains of several PACSIN-related proteins have been shown to mediate self-interactions, whereas studies using deletion mutants derived from closely related proteins led to the view that oligomerization depends on the formation of a trimeric complex via a coiled-coil region present in these molecules. To address whether the model of trimeric complex formation is applicable to PACSIN 1, the protein was recombinantly expressed and tested in four different assays for homologous interactions. The results showed that PACSIN 1 forms tetramers of about 240 kDa, with the self-interaction having a K(D) of 6.4 x 10(-8) M. Ultrastructural analysis of these oligomers after negative staining showed that laterally arranged PACSIN molecules bind to each other via a large globular domain and form a barrel-like structure. Together, these results demonstrate that the N-terminal F-BAR domain of PACSIN 1 forms the contact site for a tetrameric structure, which is able to simultaneously interact with multiple Src homology 3 binding partners.