RESUMO
Non-steroidal anti-inflammatory drugs (NSAIDs) are one of the mainstays of multimodal pain management. While effective for acute pain control, recent pre-clinical evidence has raised concerns regarding an association between NSAIDs and chronic pain and potential opioid use. Our objective was to explore the association between peri-operative use of prescription NSAIDs and the need for continued opioid prescriptions lasting 90-180 days in previously opioid-naïve patients undergoing total knee arthroplasty. A database of health claims in the USA was used to identify all opioid-naïve adult patients who underwent primary knee arthroplasty between January 2010 and October 2021. We evaluated the magnitude of association between peri-operative prescription NSAID claims and claims for opioids at 90 days postoperatively using multivariable logistic regression models. Secondary outcomes included: the magnitude of association between peri-operative NSAID prescription and claims for opioids at 180 days postoperatively; and identifying other potential factors associated with opioid claims at 90 days postoperatively. After risk adjustment using multivariable logistic regression models in the 789,736-patient cohort, the adjusted odds ratio (95%CI) for a continuous claim of opioids at 90 and 180 days postoperatively among patients with a peri-operative NSAID prescription within 30 days was 1.32 (1.30-1.35), p < 0.001; and 1.12 (1.10-1.15), p < 0.001, respectively. This estimate of effect remained robust at 90 days after accounting for known potential confounders, including pre-existing knee pain and acute postoperative pain severity. Similar analysis of other pain medications (e.g. paracetamol) did not detect such an association. This population-based cohort study suggests that peri-operative prescription NSAID use may be associated with continued opioid prescription claims at 90 and 180 days after knee arthroplasty, even after adjusting for other observed covariates for continuous opioid claims. These novel findings can inform clinical decision-making for post-surgical pain management, risk-benefit discussions with patients and future research.
Assuntos
Analgésicos Opioides , Anti-Inflamatórios não Esteroides , Artroplastia do Joelho , Dor Pós-Operatória , Humanos , Analgésicos Opioides/uso terapêutico , Anti-Inflamatórios não Esteroides/uso terapêutico , Feminino , Masculino , Estudos Retrospectivos , Dor Pós-Operatória/tratamento farmacológico , Idoso , Pessoa de Meia-Idade , Estudos de Coortes , Prescrições de Medicamentos/estatística & dados numéricos , Adulto , Assistência Perioperatória/métodosRESUMO
OBJECTIVE: Lysozyme, a 14-kDa protein, is one of the most abundant antimicrobials in the lungs. Its concentration in airway surface sufficient to kill several bacterial pathogens in vitro. The purpose of this study was to determine if administration of exogenous lysozyme would further enhance bacterial killing in vivo. METHODS: To assess the effect of acute lung infection on endogenous lysozyme protein levels, mice were infected by intratracheal instillation of Pseudomonas aeruginosa and bronchoalveolar (BAL) fluid assessed for lysozyme concentration and for muramidase activity. In order to inform in vivo testing, species-specific bacterial killing efficacy was determined by incubating mucoid P. aeruginosa with 2×105 units of chicken lysozyme, human lysozyme or with vehicle at 37°C for 2hours. Subsequently, mice challenged with intratracheally-administered mucoid P. aeruginosa, were reintubated and injected with 2×105 Units of native human lysozyme, recombinant human lysozyme or with vehicle. Lung bacterial burden was enumerated subsequently. RESULTS: The concentration of lysozyme protein in BAL fluid from mice challenged with mucoid clinical isolate of P. aeruginosa was increased 4-fold at 6hours post-infection. Quantitative culture showed that the number of recoverable bacteria was significantly decreased by both chicken and human lysozyme compared to vehicle but human lysozyme was significantly more effective than chicken egg lysozyme. In vivo, 24hours post-infection quantitative culture of lung homogenates showed that the number of viable bacteria recovered from mice treated with either native or recombinant lysozyme was decreased with 0.76±0.25×104 and 0.84±0.16×104, respectively, vs. 7.0±2.52×104 CFU/g protein in mice treated with HBSS, both P<0.05. CONCLUSIONS: These results indicate that endogenous lysozyme is increased during acute lung infection and that early administration of exogenous lysozyme further enhances bacterial killing in vivo.
Assuntos
Anti-Infecciosos/administração & dosagem , Viabilidade Microbiana/efeitos dos fármacos , Muramidase/administração & dosagem , Pneumonia Bacteriana/tratamento farmacológico , Infecções por Pseudomonas/tratamento farmacológico , Animais , Anti-Infecciosos/farmacologia , Líquido da Lavagem Broncoalveolar/microbiologia , Sinergismo Farmacológico , Quimioterapia Combinada , Feminino , Pulmão/efeitos dos fármacos , Pulmão/microbiologia , Masculino , Camundongos , Muramidase/farmacologia , Pneumonia Bacteriana/microbiologia , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/crescimento & desenvolvimento , Resultado do TratamentoRESUMO
The formation of amyloid fibrils is associated with several devastating diseases in humans and animals, including e.g. Alzheimer's disease (AD) and the spongiform encephalopathies. Here, we review and discuss the current knowledge on two amyloid peptides: lung surfactant protein C (SP-C) and the amyloid beta-peptide (Abeta), implicated in human lung disease and in AD, respectively. Both these hydrophobic peptides are derived from the transmembrane region of their precursor protein, and can transit from a monomeric alpha-helical state to a beta-sheet fibril. The alpha helices of SP-C and Abeta are composed of amino acid residues with inherently higher propensities for beta strand than helix conformation. Their helical states are stabilized by a membrane environment, and loss of membrane association thus promotes structural conversion and fibril formation. We speculate that the loss of structural context for sequences with a high propensity for formation of beta sheets may be a common feature of amyloid formation in general.
Assuntos
Peptídeos beta-Amiloides/metabolismo , Peptídeos/metabolismo , Sequência de Aminoácidos , Animais , Humanos , Neurofibrilas/metabolismo , Peptídeos/química , Peptídeos/genética , Conformação Proteica , Proteína C Associada a Surfactante PulmonarRESUMO
BACKGROUND: Improving quality of life is considered to be a major endpoint and motivation for clinical intervention in patients with perennial allergic rhinitis (PAR). In addition to classical symptoms of congestion, pruritus, and rhinorrhea, patients will often complain of not being able to sleep well at night and of feeling fatigued during the day. Like sleep apnea, PAR has also been shown to cause sleep disturbance and consequently worsen daytime fatigue and somnolence. HYPOTHESIS: It is proposed that by decreasing nasal obstruction due to allergic rhinitis by treating with the topical steroid budesonide, symptoms of daytime fatigue and somnolence can be improved. METHODS: Twenty-two subjects were enrolled in a double-blind, placebo-controlled, crossover study using Baalam's design. Patients were treated with either budesonide 128 g/day or placebo. Subjective data include the Epworth Sleepiness Scale, Functional Outcomes of Sleep Questionnaire, Rhino-conjunctivitis Quality of Life Questionnaire, and a daily diary recording nasal symptoms, sleep problems, and daytime fatigue. RESULTS: The results illustrated that the topical nasal corticosteroid significantly improved daytime fatigue (P = 0.03), somnolence (P = 0.02), and quality of sleep (P = 0.05) compared to placebo in patients suffering from PAR. SUMMARY: Budesonide is able to improve congestion, sleep, and daytime somnolence.
Assuntos
Anti-Inflamatórios/uso terapêutico , Budesonida/uso terapêutico , Distúrbios do Sono por Sonolência Excessiva/tratamento farmacológico , Distúrbios do Sono por Sonolência Excessiva/etiologia , Rinite Alérgica Perene/complicações , Rinite Alérgica Perene/tratamento farmacológico , Administração Tópica , Adulto , Estudos Cross-Over , Método Duplo-Cego , Feminino , Glucocorticoides , Humanos , Masculino , Pessoa de Meia-Idade , Obstrução Nasal/complicações , Obstrução Nasal/tratamento farmacológico , Obstrução Nasal/etiologia , Qualidade de Vida , Inquéritos e Questionários , Resultado do TratamentoRESUMO
One of the possible functions of lung surfactant protein B (SP-B), an hydrophobic membrane-associated saposin-like protein, is to reduce the alveolar surface tension by promoting insertion of phospholipids into the air/liquid interface of the lung. SP-B is a covalent homodimer; Cys48 of two polypeptides form an intermolecular disulphide bond. In order to test whether dimerisation of SP-B is important for surfactant function, transgenic mice which express (Cys48Ser) human SP-B in a mouse SP-B null background were generated. In previous studies (Cys48Ser)SP-B showed a concentration-dependent in vitro activity, suggesting that it may form non-covalent dimers. Here (Cys48Ser)SP-B isolated from bronchoalveolar lavage of transgenic mice was studied at different concentrations by circular dichroism (CD) spectroscopy, pulsating bubble surfactometry, mass spectrometry and reversed-phase HPLC. The results indicate that (Cys48Ser)SP-B, both in a phospholipid environment and in organic solvents, is largely monomeric and exhibits low activity at concentrations lower than 1 -2 microM, while at higher concentrations it forms non-covalent dimers, which are nearly functionally equivalent to native SP-B in vitro. Furthermore, electrospray mass spectrometry showed that more dimers were found relative to the monomer when the polarity of the solvent was decreased, and when the concentration of SP-B increased. (Cys48Ser)SP-B also eluted earlier than native SP-B in reversed-phase HPLC. Taken together, these results indicate that a polar surface is buried upon dimerisation, thereby promoting formation of interchain ion pairs between Glu51-Arg52' and Glu51'-Arg52.
Assuntos
Membrana Celular/efeitos dos fármacos , Proteolipídeos/farmacologia , Surfactantes Pulmonares/farmacologia , Substituição de Aminoácidos , Animais , Dicroísmo Circular , Dimerização , Relação Dose-Resposta a Droga , Membranas Artificiais , Camundongos , Camundongos Transgênicos , Estrutura Secundária de Proteína , Proteolipídeos/química , Proteolipídeos/genética , Surfactantes Pulmonares/química , Surfactantes Pulmonares/genética , Tensão Superficial/efeitos dos fármacosRESUMO
Proteolytic processing of surfactant protein C (SP-C) proprotein in multivesicular bodies of alveolar type II cells results in a 35-residue mature peptide, consisting of a transmembrane domain and a 10-residue extramembrane domain. SP-C mature peptide is stored in lamellar bodies (a lysosomal-like organelle) and secreted with surfactant phospholipids into the alveolar space. This study was designed to identify the peptide domain of SP-C required for sorting and secretion of this integral membrane peptide. Deletion analyses in transiently transfected PC12 cells and isolated mouse type II cells suggested the extramembrane domain of mature SP-C was cytosolic and sufficient for sorting to the regulated secretory pathway. Intratracheal injection of adenovirus encoding SP-C mature peptide resulted in secretion into the alveolar space of wild type mice but not SP-C (-/-) mice. SP-C secretion in null mice was restored by the addition of the N-terminal propeptide. The cytosolic domain, consisting of the N- terminal propeptide and extramembrane domain of mature SP-C peptide, supported secretion of the transmembrane domain of platelet-derived growth factor receptor. Collectively, these studies indicate that the N-terminal propeptide of SP-C is required for intracellular sorting and secretion of SP-C.
Assuntos
Proteínas de Membrana/metabolismo , Proteolipídeos/metabolismo , Surfactantes Pulmonares/metabolismo , Sequência de Aminoácidos , Animais , Endocitose , Humanos , Proteínas de Membrana/química , Células PC12 , Proteolipídeos/química , Surfactantes Pulmonares/química , RatosRESUMO
SP-B is the only surfactant-associated protein absolutely required for postnatal lung function and survival. Complete deficiency of SP-B in mice and humans results in lethal, neonatal respiratory distress syndrome and is characterized by a virtual absence of lung compliance, highly disorganized lamellar bodies, and greatly diminished levels of SP-C mature peptide; in contrast, lung structure and function in SP-C null mice is normal. This review attempts to integrate recent findings in humans and transgenic mice with the results of in vitro studies to provide a better understanding of the functions of SP-B and SP-C and the structural basis for their actions.
Assuntos
Proteolipídeos/genética , Proteolipídeos/metabolismo , Alvéolos Pulmonares/fisiologia , Surfactantes Pulmonares/genética , Surfactantes Pulmonares/metabolismo , Sequência de Aminoácidos , Animais , Humanos , Dados de Sequência MolecularRESUMO
Allergen challenge in the lung of humans and animals is associated with surfactant dysfunction, but the mechanism of this effect has not been established. By using a murine model of asthma we now report the effect of allergen-induced airway inflammation on the expression of transgenes regulated by the human surfactant protein (hSP)-C promoter. The hSP-C 3.7-kilobase pair promoter was used to direct the expression of eotaxin, an eosinophil-selective chemokine, into the lungs of several transgenic lines. As expected, the transgenic mice expressed increased amounts of eotaxin mRNA and protein compared with wild-type mice. Surprisingly, following allergen challenge, there was a marked down-regulation of transgene mRNA in three independent transgenic lines. The down-regulation was in contrast to other related proteins such as endogenous eotaxin and surfactant protein D levels, which were both increased following allergen challenge. Consistent with specific down-regulation of the eotaxin transgene, there was no increase in pulmonary eosinophil levels in the transgenic mice above that found in wild-type mice. Analysis of hSP-C transgenic mice with distinct reporter genes and 3'-untranslated regions revealed that allergen challenge was directly affecting the hSP-C promoter. We hypothesized that allergen-induced down-regulation of the hSP-C promoter was related to the eosinophilic inflammation. To test this, we blocked eosinophilic inflammation in the lungs by treating mice with neutralizing antiserum against interleukin-5. Interestingly, this treatment also blocked allergen-induced inhibition of the hSP-C promoter. These results establish that allergic airway inflammation is associated with up-regulation of the surfactant proteins primarily involved in immunity, whereas down-regulation of the surfactant protein primarily involved in maintaining airway patency. Furthermore, the marked down-regulation of the hSP-C promoter is interleukin-5-dependent, implying a critical role for eosinophilic inflammation. These results suggest that alterations in surfactant protein levels may contribute to immune and airway dysfunction in asthma.
Assuntos
Asma/metabolismo , Quimiocinas CC , Citocinas/fisiologia , Interleucina-5/fisiologia , Regiões Promotoras Genéticas , Proteolipídeos/genética , Surfactantes Pulmonares/genética , Regiões 3' não Traduzidas/fisiologia , Animais , Asma/etiologia , Quimiocina CCL11 , Regulação para Baixo , Camundongos , Camundongos Transgênicos , Proteolipídeos/fisiologia , Surfactantes Pulmonares/fisiologia , TransgenesRESUMO
Neurobehavioral performance and alterations in mood consequent to sleep disorders and their treatment has recently been the focus of clinical investigations. Primarily in patients with obstructive sleep apnea (OSA), there has been increased interest in the effects of sleep fragmentation and hypoxemia on the brain's ability to process information and to alter affect. The purpose of the second part of this two-part series is to describe measures that could be applied to document the impact of sleep disorders on neurobehavioral performance and mood, discuss factors affecting the selection of measures for research and practice, and to describe evidence generated by the use of these instruments in research. The neurobehavioral deficits that accompany sleep deprivation can be categorised as decrements in cognitive throughput, working memory and sustained attention. Usually evaluated using tasks of short duration, impairments associated with OSA have included impaired information processing and decline in the total number of completed and/or correct responses per unit time. Using assessments of working memory, including short recall maneuvers involving words or paragraphs, investigators have documented in sleep apnea patients the inability to consolidate and recall material producing deficits in the recollection and retention of new information. Evaluations of sustained attention appraise reaction time, the ability to remain on task, and the number of errors of omission and false responses. Evidence suggests that OSA patients display slowing of response time as well as increased errors, lapses and number of false responses. Similar deficits have been documented with sustained and divided attention tasks that present the respondent with challenges in tracking and reaction response, tasks required for driving. Although untreated sleep apnea patients were more impaired than normal controls, their performance on such tasks was greatly enhanced following CPAP treatment. In addition to substantial clinically meaningful improvements in many areas of neurobehavioral performance, changes in mood have also accompanied treatment for sleep apnea. 2001 Harcourt Publishers Ltd
RESUMO
To assess the role of lysozyme in pulmonary host defense in vivo, transgenic mice expressing rat lysozyme cDNA in distal respiratory epithelial cells were generated. Two transgenic mouse lines were established in which the level of lysozyme protein in bronchoalveolar (BAL) lavage fluid was increased 2- or 4-fold relative to that in WT mice. Lung structure and cellular composition of BAL were not altered by the expression of lysozyme. Lysozyme activity in BAL was significantly increased (6.6- and 17-fold) in 5-wk-old animals from each transgenic line. To determine whether killing of bacteria was enhanced by expression of rat lysozyme, 5-wk-old transgenic mice and WT littermates were infected with 10(6) CFU of group B streptococci or 10(7) CFU of a mucoid strain of Pseudomonas aeruginosa by intratracheal injection. Killing of group B streptococci was significantly enhanced (2- and 3-fold) in the mouse transgenic lines at 6 h postinfection and was accompanied by a decrease in systemic dissemination of pathogen. Killing of Pseudomonas aeruginosa was also enhanced in the transgenic lines (5- and 30-fold). Twenty-four hours after administration of Pseudomonas aeruginosa, all transgenic mice survived, whereas 20% of the WT mice died. Increased production of lysozyme in respiratory epithelial cells of transgenic mice enhanced bacterial killing in the lung in vivo, and was associated with decreased systemic dissemination of pathogen and increased survival following infection.
Assuntos
Adjuvantes Imunológicos/biossíntese , Adjuvantes Imunológicos/genética , Pulmão/enzimologia , Pulmão/microbiologia , Muramidase/biossíntese , Muramidase/genética , Pseudomonas aeruginosa/crescimento & desenvolvimento , Pseudomonas aeruginosa/imunologia , Adjuvantes Imunológicos/fisiologia , Animais , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Movimento Celular/genética , Movimento Celular/imunologia , Regulação da Expressão Gênica/imunologia , Humanos , Mediadores da Inflamação/análise , Pulmão/imunologia , Camundongos , Camundongos Endogâmicos , Camundongos Transgênicos , Muramidase/fisiologia , Neutrófilos/imunologia , Neutrófilos/patologia , Infecções por Pseudomonas/enzimologia , Infecções por Pseudomonas/genética , Infecções por Pseudomonas/imunologia , Infecções por Pseudomonas/microbiologia , Ratos , Transgenes/imunologiaRESUMO
Surfactant protein B (SP-B) -/- mice die of lethal respiratory distress syndrome shortly after birth. Alveolar type II epithelial cells in SP-B-deficient mice are characterized by a complete absence of lamellar bodies, the intracellular storage form of pulmonary surfactant, and the presence of inclusions containing numerous small vesicles and electron-dense masses. The present study was undertaken to characterize the formation of these inclusions during fetal lung development and clarify their relationship to lamellar bodies. In wild-type and SP-B +/- mice, small lamellar bodies with loosely organized lamellae and distinct limiting membranes were first detected on day 16 to 16.5 of gestation. SP-B -/- mice were readily identified on day 16 by the absence of immature lamellar bodies, the appearance of vesicular inclusions similar to those previously described in late gestation SP-B -/- mice, and the accumulation of misprocessed SP-C protein. Vesicular inclusions were rarely detected in SP-B +/- mice and were never detected in wild-type littermates. Classical multivesicular bodies were observed fusing with lamellar bodies in wild-type mice, and with the vesicular inclusions in SP-B -/- mice that occasionally contained a few membrane lamellae. On day 18, the airways of SP-B -/- mice lacked tubular myelin and were filled with vesicles and electron-dense masses, suggesting that the contents of the vesicular inclusions were secreted. Taken together, these observations suggest that vesicular inclusions in SP-B -/- mice are disorganized lamellar bodies in which the absence of SP-B leads to failure to package surfactant phospholipids into concentric lamellae.
Assuntos
Pulmão/embriologia , Proteolipídeos/genética , Surfactantes Pulmonares/genética , Animais , Pulmão/metabolismo , Pulmão/patologia , Camundongos , Camundongos KnockoutRESUMO
Surfactant protein B (SP-B) is detected in the airways as a sulfhydryl-dependent dimer (M(r) approximately 16,000). To test the hypothesis that formation of homodimers is critical for SP-B function, the cysteine residue reported to be involved in SP-B dimerization was mutated to serine (Cys(248) --> Ser) and the mutated protein was targeted to the distal respiratory epithelium of transgenic mice. Transgenic lines which demonstrated appropriate processing, sorting, and secretion of human SP-B monomer were crossed with SP-B +/- mice to achieve expression of human monomer in the absence of endogenous SP-B dimer (hSP-B(mon), mSP-B-/-). In two of three transgenic lines, hSP-B(mon), mSP-B-/- mice had normal lung structure, complete processing of SP-C proprotein, well formed lamellar bodies, and normal longevity. Pulmonary function studies revealed an altered hysteresis curve for hSP-B(mon), mSP-B-/- mice relative to wild type mice. Large aggregate surfactant fractions from hSP-B(mon), mSP-B-/- mice resulted in higher minimum surface tension in vitro compared with surfactant from wild type mice. Surfactant lipids supplemented with 2% hSP-B monomer resulted in slower adsorption and higher surface tension than surfactant with 2% hSP-B dimer. Taken together, these data indicate a role for SP-B dimer in surface tension reduction in the alveolus.
Assuntos
Apoproteínas/metabolismo , Proteínas Associadas a Surfactantes Pulmonares , Surfactantes Pulmonares/metabolismo , Animais , Apoproteínas/química , Apoproteínas/genética , Dimerização , Humanos , Camundongos , Camundongos Transgênicos , Mutação , Surfactantes Pulmonares/química , Surfactantes Pulmonares/genética , Relação Estrutura-AtividadeRESUMO
The 79-amino acid, mature SP-B peptide contains three intramolecular disulfide bonds shared by all saposin-like proteins. This study tested the hypothesis that the disulfide bond formed between cysteine residues 35 and 46 (residues 235 and 246 of the SP-B proprotein) is essential for proper function of SP-B. To test the role of this bridge in SP-B function in vivo, a construct was generated in which cysteine residues 235 and 246 of the human SP-B proprotein were mutated to serine and cloned under the control of the 3.7-kilobase hSP-C promoter (hSP-B(C235S/C246S)). In two transgenic mouse lines, expression of the mutant peptide in the wild-type murine SP-B background was invariably lethal in the neonatal period. In four additional lines, survival was inversely related to the level of transgene expression. To test the ability of the mutant peptide to functionally replace the wild-type protein, transgenic mice were crossed into the SP-B null background. No animals that expressed hSP-B(C235S/C246S) in the murine SP-B-/- background survived the neonatal period. hSP-B(C235S/C246S) proprotein accumulated in the endoplasmic reticulum and was not processed to the mature, biologically active peptide. The results of these studies demonstrate that the intramolecular bridge between residues 235 and 246 is critical for intracellular trafficking of SP-B and suggest that overexpression of mutant SP-B in the wild-type background may be lethal.
Assuntos
Apoproteínas/química , Proteínas Associadas a Surfactantes Pulmonares , Surfactantes Pulmonares/química , Animais , Apoproteínas/genética , Apoproteínas/metabolismo , Dissulfetos , Humanos , Camundongos , Camundongos Transgênicos , Mutação , Surfactantes Pulmonares/genética , Surfactantes Pulmonares/metabolismo , Relação Estrutura-AtividadeRESUMO
To determine the role of surfactant protein B (SP-B) in bacterial clearance from the airways, three groups of mice expressing different levels of SP-B were studied: wild-type mice, hemizygous SP-B mice, and SP-B overexpressing transgenic mice. SP-B levels in overexpressing mice were increased 5-fold relative to hemizygous mice and 2- to 3-fold over wild-type littermates. Mice from each group were infected intratracheally with the common airway pathogens, group B streptococci or Pseudomonas aeruginosa. There was no significant difference in the number of recoverable viable bacteria at 6 h (group B streptococci and P. aeruginosa) and at 24 h (P. aeruginosa) among the three groups. Similarly, systemic dissemination of bacteria was not different among the three groups for both pathogens and at both time points. We conclude that SP-B levels in vivo do not influence clearance of bacteria from the lungs.
Assuntos
Pulmão/microbiologia , Proteolipídeos/metabolismo , Pseudomonas aeruginosa/metabolismo , Surfactantes Pulmonares/metabolismo , Streptococcus agalactiae/metabolismo , Animais , Taxa de Depuração Metabólica , Camundongos , Camundongos Transgênicos , Solubilidade , Água/químicaRESUMO
Pulmonary surfactant protein B (SP-B) is a 79 amino acid peptide that is intimately associated with surfactant phospholipids in the alveolar airspace. Mutations of the SP-B gene that result in complete absence of SP-B are invariably fatal in the neonatal period. The pathology associated with SP-B deficiency suggests that SP-B plays a critical role in integrating the synthesis, assembly and metabolism of the surfactant complex. A strategy is described to elucidate the role of SP-B in surfactant homeostasis by characterizing the pathophysiology associated with cell specific expression of SP-B constructs in vivo. Human SP-B constructs, under control of lung cell-specific promoters, were expressed in SP-B knockout mice in order to achieve expression of the human transgene in a null background. The effect of transgene expression on lung structure and function was assessed by biochemical, morphological and physiological analyses of the surfactant system in fetal and postnatal offspring.
Assuntos
Camundongos Knockout , Proteolipídeos/química , Proteolipídeos/fisiologia , Surfactantes Pulmonares/química , Surfactantes Pulmonares/fisiologia , Animais , Dimerização , Humanos , Pulmão/fisiologia , Camundongos , Camundongos Transgênicos , Mutagênese , Proteolipídeos/genética , Surfactantes Pulmonares/genéticaRESUMO
Surfactant protein B (SP-B) mRNA and protein are restricted to alveolar Type II and Clara cells in the respiratory epithelium. In order to investigate the function of SP-B in these distinct cell types, transgenic mice were generated in which SP-B expression was selectively restored in Type II cells or Clara cells of SP-B -/- mice. The 4.8-kilobase murine SP-C promoter was used to generate 3 transgenic lines which expressed human SP-B in Type II cells (mSP-C/hSP-B). Likewise, the 2.3-kilobase murine CCSP promoter was used to generate two transgenic lines which expressed human SP-B in Clara cells (mCCSP/hSP-B). mSP-C/hSP-B and mCCSP/hSP-B transgenic mice were subsequently bred to SP-B +/- mice in order to selectively express SP-B in Type II cells or Clara cells of SP-B -/- mice. Selective restoration of SP-B expression in Type II cells completely rescued the neonatal lethal phenotype in SP-B -/- mice. Expression of SP-B in some, but not all Type II cells of SP-B -/- mice, allowed postnatal survival, but resulted in significantly altered lung architecture and function. Selective restoration of SP-B expression in Clara cells of SP-B -/- mice resulted in respiratory dysfunction and invariable neonatal death, related to the complete absence of mature SP-B peptide in these mice. These results indicate that expression and processing of the SP-B proprotein to the mature peptide in Type II cells is absolutely required for lung function in vivo and that SP-B expression in Clara cells cannot substitute for this function.
Assuntos
Brônquios/metabolismo , Proteolipídeos/biossíntese , Alvéolos Pulmonares/metabolismo , Surfactantes Pulmonares/biossíntese , Animais , Western Blotting , Brônquios/citologia , Células Epiteliais/metabolismo , Humanos , Hibridização In Situ , Camundongos , Camundongos Transgênicos , Fenótipo , Proteolipídeos/genética , Surfactantes Pulmonares/genéticaRESUMO
Two small, hydrophobic peptides, surfactant protein (SP)-B and SP-C, play important roles in the generation and maintenance of a surface active film in the alveolus. Isolation and characterization of the cDNAs encoding SP-B and SP-C indicate that both peptides are synthesized as larger proproteins which are proteolytically processed to peptides with Mr approx. 8000 and 4000, respectively. The biosynthetic pathway leading to generation and secretion of the biophysically active mature SP-B and SP-C peptides is reviewed.
Assuntos
Processamento de Proteína Pós-Traducional , Proteolipídeos/metabolismo , Surfactantes Pulmonares/metabolismo , Sequência de Aminoácidos , Transporte Biológico , Brônquios/citologia , Brônquios/metabolismo , Modelos Moleculares , Dados de Sequência Molecular , Alvéolos Pulmonares/citologia , Alvéolos Pulmonares/metabolismoRESUMO
BACKGROUND: This article describes the development and psychometric evaluation of the Pulmonary Functional Status Scale (PFSS). The PFSS was developed to address the need for a self-administered, disease-specific, functional status outcome measure that was conceptually based, relatively short, and comprehensively assessed the psychosocial domains affected by chronic pulmonary disease. METHODS: Content validity of the PFSS was established by a panel of experts. Construct validity was determined by principal components factor analysis using data from 365 subjects derived from several studies that used the PFSS. The sample was predominantly white (80%), married (36%), and male (56%) with a mean age of 68.1 years, mean forced expiratory volume in one second of 1.18 L and mean forced vital capacity of 2.31 L. Test-retest reliability was evaluated in 17 outpatients with chronic obstructive pulmonary disease. Concurrent validity was assessed by correlating responses on the total PFSS with the Sickness Impact Profile (SIP) score (n = 39), and the 12-minute walk test (n = 104). RESULTS: The factor structure solution used 35 of the 50 submitted items, explained 48.7% of the variance, and yielded three factors: Daily Activities/Social Functioning, Psychological Functioning, and Sexual Functioning. Cronbach's alpha coefficients for the total PFSS was 0.93 and the test-retest correlation coefficient was rho = 0.75 (P = 0.001). The correlation between the total PFSS score and the SIP and 12-minute walk test were r = -0.54 (P < 0.001) and r = 0.62 (P < 0.001), respectively. CONCLUSION: The PFSS has solid psychometric properties that make it acceptable for use in clinical practice as well as research.
Assuntos
Pneumopatias Obstrutivas/psicologia , Qualidade de Vida , Idoso , Doença Crônica , Feminino , Humanos , Pneumopatias Obstrutivas/fisiopatologia , Pneumopatias Obstrutivas/reabilitação , Masculino , Avaliação de Resultados em Cuidados de Saúde , Psicometria , Reprodutibilidade dos Testes , Respiração/fisiologia , Testes de Função Respiratória , Estudos Retrospectivos , Inquéritos e QuestionáriosRESUMO
Pulmonary function was assessed in newborn wild-type and homozygous and heterozygous surfactant protein B (SP-B)-deficient mice after birth. SP-B +/+ and SP-B+/- mice became well oxygenated and survived postnatally. Although lung compliance was decreased slightly in the SP-B+/- mice, lung volumes and compliances were decreased markedly in homozygous SP-B-/- mice. They died rapidly after birth, failing to inflate their lungs or oxygenate. SP-B proprotein was absent in the SP-B-/- mice and was reduced in the SP-B+/- mice, as assessed by Western analysis. Surfactant protein A, surfactant proprotein C, surfactant protein D, and surfactant phospholipid content in lungs from SP-B+/- and SP-B-/- mice were not altered. Lung saturated phosphatidylcholine and precursor incorporation into saturated phosphatidylcholine were not influenced by SP-B genotype. Intratracheal administration of perfluorocarbon resulted in lung expansion, oxygenation, and prolonged survival of SP-B-/- mice and in reduced lung compliance in SP-B+/+ and SP-B+/- mice. Lack of SP-B caused respiratory failure at birth, and decreased SP-B protein was associated with reduced lung compliance. These findings demonstrate the critical role of SP-B in perinatal adaptation to air breathing.
Assuntos
Pulmão/fisiologia , Pulmão/fisiopatologia , Proteolipídeos/genética , Proteolipídeos/metabolismo , Surfactantes Pulmonares/genética , Surfactantes Pulmonares/metabolismo , Animais , Animais Recém-Nascidos , Fluorocarbonos/toxicidade , Heterozigoto , Homozigoto , Pulmão/patologia , Camundongos , Camundongos Knockout , Proteolipídeos/biossíntese , Proteínas Associadas a Surfactantes Pulmonares , Surfactantes Pulmonares/biossínteseRESUMO
Glycogen accumulates to significant levels in epithelial cells of the developing respiratory tract. Mobilization of glycogen stores is regulated differentially along the respiratory epithelium such that glycogenolysis in the alveolar epithelium (the site of surfactant synthesis) precedes that in the bronchial and bronchiolar epithelium. The initial step in glycogen degradation is catalyzed by glycogen phosphorylase, which exists as three genetically distinct isozymes referred to as muscle, liver, and brain isoforms. The goal of this study was to characterize the temporal and spatial expression of each of the glycogen phosphorylase isozymes in developing lung to determine which isoform(s) was associated with glycogen mobilization in the fetal type II epithelial cell. RNA levels encoding glycogen phosphorylase were assessed by ribonuclease protection assay using isoform-specific antisense probes. RNAs encoding the brain and liver isozymes were detected in isolated day 20 fetal type II epithelial cells and at lower levels in adult type II cells. The muscle isoform RNA was barely detectable in fetal type II cells and was undetectable in adult type II cells. Expression of brain and liver isoform RNAs was higher in whole fetal lung than in fetal type II cells. Consistent with this result, in situ hybridization studies demonstrated widespread expression of the brain and liver isoforms in developing lung tissues; in contrast, expression of the muscle isoform was restricted to the pulmonary vein. Glycogen phosphorylase enzyme activity corresponding to the brain isoform was clearly detected in isolated fetal type II cells; however, the majority of enzyme activity migrated as two bands with distinct electrophoretic mobilities that may have been the result of isoform heterodimerization. Collectively, these results suggest that the brain and liver isoforms of glycogen phosphorylase may be involved in mobilization of type II cell glycogen during late fetal lung development.