RESUMO
BACKGROUND: It is estimated that 1-13% of cases of bronchiectasis in adults globally are attributable to primary ciliary dyskinesia (PCD) but many adult patients with bronchiectasis have not been investigated for PCD. PCD is a disorder caused by mutations in genes required for motile cilium structure or function, resulting in impaired mucociliary clearance. Symptoms appear in infancy but diagnosis is often late or missed, often due to the lack of a "gold standard" diagnostic tool and non-specific symptoms. Mutations in > 50 genes account for around 70% of cases, with additional genes, and non-coding, synonymous, missense changes or structural variants (SVs) in known genes presumed to account for the missing heritability. METHODS: UK patients with no identified genetic confirmation for the cause of their PCD or bronchiectasis were eligible for whole genome sequencing (WGS) in the Genomics England Ltd 100,000 Genomes Project. 21 PCD probands and 52 non-cystic fibrosis (CF) bronchiectasis probands were recruited in Wessex Genome Medicine Centre (GMC). We carried out analysis of single nucleotide variants (SNVs) and SVs in all families recruited in Wessex GMC. RESULTS: 16/21 probands in the PCD cohort received confirmed (n = 9), probable (n = 4) or possible (n = 3) diagnosis from WGS, although 13/16 of these could have been picked up by current standard of care gene panel testing. In the other cases, SVs were identified which were missed by panel testing. We identified variants in novel PCD candidate genes (IFT140 and PLK4) in 2 probands in the PCD cohort. 3/52 probands in the non-CF bronchiectasis cohort received a confirmed (n = 2) or possible (n = 1) diagnosis of PCD. We identified variants in novel PCD candidate genes (CFAP53 and CEP164) in 2 further probands in the non-CF bronchiectasis cohort. CONCLUSIONS: Genetic testing is an important component of diagnosing PCD, especially in cases of atypical disease history. WGS is effective in cases where prior gene panel testing has found no variants or only heterozygous variants. In these cases it may detect SVs and is a powerful tool for novel gene discovery.
Assuntos
Transtornos da Motilidade CiliarRESUMO
Primary ciliary dyskinesia (PCD) is a rare disease with autosomal recessive inheritance, caused mostly by bi-allelic gene mutations that impair motile cilia structure and function. Currently, there are no causal treatments for PCD. In many disease models, translational readthrough of premature termination codons (PTC-readthrough) induced by aminoglycosides has been proposed as an effective way of restoring functional protein expression and reducing disease symptoms. However, variable outcomes of pre-clinical trials and toxicity associated with long-term use of aminoglycosides prompt the search for other compounds that might overcome these problems. Because a high proportion of PCD-causing variants are nonsense mutations, readthrough therapies are an attractive option. We tested a group of chemical compounds with known PTC-readthrough potential (ataluren, azithromycin, tylosin, amlexanox, and the experimental compound TC007), collectively referred to as non-aminoglycosides (NAGs). We investigated their PTC-readthrough efficiency in six PTC mutations found in Polish PCD patients, in the context of cell and cilia health, and in comparison to the previously tested aminoglycosides. The NAGs did not compromise the viability of the primary nasal respiratory epithelial cells, and the ciliary beat frequency was retained, similar to what was observed for gentamicin. In HEK293 cells transfected with six PTC-containing inserts, the tested compounds stimulated PTC-readthrough but with lower efficiency than aminoglycosides. The study allowed us to select compounds with minimal negative impact on cell viability and function but still the potential to induce PTC-readthrough.
Assuntos
Aminoglicosídeos/farmacologia , Transtornos da Motilidade Ciliar/genética , Códon sem Sentido/genética , Mutação/genética , Biossíntese de Proteínas/genética , Morte Celular/efeitos dos fármacos , Células Cultivadas , Cílios/efeitos dos fármacos , Cílios/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células HEK293 , Humanos , Nariz/patologia , Biossíntese de Proteínas/efeitos dos fármacosRESUMO
Angiotensin-converting enzyme 2 (ACE2) is the main entry point in airway epithelial cells for SARS-CoV-2. ACE2 binding to the SARS-CoV-2 protein spike triggers viral fusion with the cell plasma membrane, resulting in viral RNA genome delivery into the host. Despite ACE2's critical role in SARS-CoV-2 infection, full understanding of ACE2 expression, including in response to viral infection, remains unclear. ACE2 was thought to encode five transcripts and one protein of 805 amino acids. In the present study, we identify a novel short isoform of ACE2 expressed in the airway epithelium, the main site of SARS-CoV-2 infection. Short ACE2 is substantially upregulated in response to interferon stimulation and rhinovirus infection, but not SARS-CoV-2 infection. This short isoform lacks SARS-CoV-2 spike high-affinity binding sites and, altogether, our data are consistent with a model where short ACE2 is unlikely to directly contribute to host susceptibility to SARS-CoV-2 infection.
Assuntos
Enzima de Conversão de Angiotensina 2/genética , COVID-19/genética , Células Epiteliais/metabolismo , Animais , Sítios de Ligação , Células Cultivadas , Chlorocebus aethiops , Éxons , Células HEK293 , Humanos , Interferons/imunologia , Ligação Proteica , Isoformas de Proteínas/genética , Sítios de Splice de RNA , RNA-Seq , Sistema Respiratório/citologia , Glicoproteína da Espícula de Coronavírus/metabolismo , Transcriptoma , Regulação para Cima , Células VeroRESUMO
Non-typeable Haemophilus influenzae (NTHi) is the most common pathogen in primary ciliary dyskinesia (PCD) patients. We hypothesised that abnormal ciliary motility and low airway nitric oxide (NO) levels on airway epithelial cells from PCD patients might be permissive for NTHi colonisation and biofilm development.We used a primary epithelial cell co-culture model to investigate NTHi infection. Primary airway epithelial cells from PCD and non-PCD patients were differentiated to ciliation using an air-liquid interface culture and then co-cultured with NTHi.NTHi adherence was greater on PCD epithelial cells compared to non-PCD cells (p<0.05) and the distribution of NTHi on PCD epithelium showed more aggregated NTHi in biofilms (p<0.001). Apart from defective ciliary motility, PCD cells did not significantly differ from non-PCD epithelial cells in the degree of ciliation and epithelial integrity or in cytokine, LL-37 and NO production. Treatment of PCD epithelia using exogenous NO and antibiotic significantly reduced NTHi viability in biofilms compared with antibiotic treatment alone.Impaired ciliary function was the primary defect in PCD airway epithelium underlying susceptibility to NTHi biofilm development compared with non-PCD epithelium. Although NO responses were similar, use of targeted NO with antibiotics enhanced killing of NTHi in biofilms, suggesting a novel therapeutic approach.
Assuntos
Células Epiteliais/microbiologia , Infecções por Haemophilus/fisiopatologia , Síndrome de Kartagener/microbiologia , Óxido Nítrico/farmacologia , Adolescente , Adulto , Antibacterianos/farmacologia , Aderência Bacteriana , Proteínas de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Estudos de Casos e Controles , Criança , Pré-Escolar , Citocinas/metabolismo , Feminino , Haemophilus influenzae/patogenicidade , Haemophilus influenzae/fisiologia , Humanos , Síndrome de Kartagener/fisiopatologia , Masculino , Pessoa de Meia-Idade , Cultura Primária de Células , Adulto JovemRESUMO
High levels of Pim expression have been implicated in several hematopoietic and solid tumor cancers, suggesting that inhibition of Pim signaling could provide patients with therapeutic benefit. Herein, we describe our progress towards this goal using a screening hit (rac-1) as a starting point. Modification of the indazole ring resulted in the discovery of a series of imidazopyridazine-based Pim inhibitors exemplified by compound 22m, which was found to be a subnanomolar inhibitor of the Pim-1 and Pim-2 isoforms (IC50 values of 0.024nM and 0.095nM, respectively) and to potently inhibit the phosphorylation of BAD in a cell line that expresses high levels of all Pim isoforms, KMS-12-BM (IC50=28nM). Profiling of Pim-1 and Pim-2 expression levels in a panel of multiple myeloma cell lines and correlation of these data with the potency of compound 22m in a proliferation assay suggests that Pim-2 inhibition would be advantageous for this indication.
Assuntos
Inibidores de Proteínas Quinases/química , Inibidores de Proteínas Quinases/farmacologia , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-pim-1/antagonistas & inibidores , Proteínas Proto-Oncogênicas/antagonistas & inibidores , Piridazinas/química , Piridazinas/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Humanos , Modelos Moleculares , Mieloma Múltiplo/tratamento farmacológico , Mieloma Múltiplo/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-pim-1/metabolismo , Relação Estrutura-AtividadeRESUMO
Nasal drug administration is a promising alternative to oral and parenteral administration for both local and systemic delivery of drugs. The benefits include its noninvasive nature, rapid absorption, and circumvention of first pass metabolism. Hence, the use of an in vitro model using human primary nasal epithelial cells could be key to understanding important functions and parameters of the respiratory epithelium. This model will enable investigators to address important and original research questions using a biologically relevant in vitro platform that mimics the in vivo nasal epithelial physiology. The purpose of this study was to establish, systematically characterize, and validate the use of a primary human nasal epithelium model cultured at the air-liquid interface for the study of inflammatory responses and drug transport and to simultaneously quantify drug effects on ciliary activity.
Assuntos
Células Epiteliais/fisiologia , Mucosa Nasal/fisiologia , Preparações Farmacêuticas/administração & dosagem , Mucosa Respiratória/fisiologia , Administração Intranasal/métodos , Adulto , Técnicas de Cultura de Células/métodos , Células Cultivadas , Sistemas de Liberação de Medicamentos/métodos , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Airway NO synthase (NOS) isoenzymes are responsible for rapid and localised nitric oxide (NO) production and are expressed in airway epithelium. We sought to determine the localisation of neuronal NOS (nNOS) in airway epithelium due to the paucity of evidence. METHODS AND RESULTS: Sections of healthy human bronchial tissue in glycol methacrylate resin and human nasal polyps in paraffin wax were immunohistochemically labelled and reproducibly demonstrated nNOS immunoreactivity, particularly at the proximal portion of cilia; this immunoreactivity was blocked by a specific nNOS peptide fragment. Healthy human epithelial cells differentiated at an air-liquid interface (ALI) confirmed the presence of all three NOS isoenzymes by immunofluorescence labelling. Only nNOS immunoreactivity was specific to the ciliary axonemeand co-localised with the cilia marker ß-tubulin in the proximal part of the ciliary axoneme. CONCLUSIONS: We report a novel localisation of nNOS at the proximal portion of cilia in airway epithelium and conclude that its independent and local regulation of NO levels is crucial for normal cilia function.
Assuntos
Cílios/enzimologia , Óxido Nítrico Sintase Tipo I/metabolismo , Mucosa Respiratória/enzimologia , Brônquios/química , Brônquios/enzimologia , Células Cultivadas , Cílios/química , Cílios/metabolismo , Humanos , Imuno-Histoquímica , Pólipos Nasais/química , Pólipos Nasais/enzimologia , Óxido Nítrico Sintase Tipo I/química , Mucosa Respiratória/química , Mucosa Respiratória/citologia , Mucosa Respiratória/metabolismoRESUMO
BACKGROUND: The diagnosis of primary ciliary dyskinesia (PCD) requires the analysis of ciliary function and ultrastructure. Diagnosis can be complicated by secondary effects on cilia such as damage during sampling, local inflammation or recent infection. To differentiate primary from secondary abnormalities, re-analysis of cilia following culture and re-differentiation of epithelial cells at an air-liquid interface (ALI) aids the diagnosis of PCD. However changes in ciliary beat pattern of cilia following epithelial cell culture has previously been described, which has brought the robustness of this method into question. This is the first systematic study to evaluate ALI culture as an aid to diagnosis of PCD in the light of these concerns. METHODS: We retrospectively studied changes associated with ALI-culture in 158 subjects referred for diagnostic testing at two PCD centres. Ciliated nasal epithelium (PCD n = 54; non-PCD nâ 111) was analysed by high-speed digital video microscopy and transmission electron microscopy before and after culture. RESULTS: Ciliary function was abnormal before and after culture in all subjects with PCD; 21 PCD subjects had a combination of static and uncoordinated twitching cilia, which became completely static following culture, a further 9 demonstrated a decreased ciliary beat frequency after culture. In subjects without PCD, secondary ciliary dyskinesia was reduced. CONCLUSIONS: The change to ciliary phenotype in PCD samples following cell culture does not affect the diagnosis, and in certain cases can assist the ability to identify PCD cilia.
Assuntos
Cílios/genética , Transtornos da Motilidade Ciliar/genética , Síndrome de Kartagener/genética , Ar , Técnicas de Cultura de Células , Células Cultivadas , Cílios/fisiologia , Transtornos da Motilidade Ciliar/diagnóstico , Células Epiteliais/citologia , Humanos , Síndrome de Kartagener/diagnóstico , Microscopia Eletrônica de Transmissão , Microscopia de Vídeo , Mucosa Nasal , Fenótipo , Estudos RetrospectivosRESUMO
The phosphoinositide 3-kinase family catalyzes the phosphorylation of phosphatidylinositol-4,5-diphosphate to phosphatidylinositol-3,4,5-triphosphate, a secondary messenger which plays a critical role in important cellular functions such as metabolism, cell growth, and cell survival. Our efforts to identify potent, efficacious, and orally available phosphatidylinositol 3-kinase (PI3K) inhibitors as potential cancer therapeutics have resulted in the discovery of 4-(2-((6-methoxypyridin-3-yl)amino)-5-((4-(methylsulfonyl)piperazin-1-yl)methyl)pyridin-3-yl)-6-methyl-1,3,5-triazin-2-amine (1). In this paper, we describe the optimization of compound 1, which led to the design and synthesis of pyridyltriazine 31, a potent pan inhibitor of class I PI3Ks with a superior pharmacokinetic profile. Compound 31 was shown to potently block the targeted PI3K pathway in a mouse liver pharmacodynamic model and inhibit tumor growth in a U87 malignant glioma glioblastoma xenograft model. On the basis of its excellent in vivo efficacy and pharmacokinetic profile, compound 31 was selected for further evaluation as a clinical candidate and was designated AMG 511.
Assuntos
Inibidores de Fosfoinositídeo-3 Quinase , Inibidores de Proteínas Quinases/farmacologia , Triazinas/farmacologia , Cristalografia por Raios X , Modelos Moleculares , Inibidores de Proteínas Quinases/químicaRESUMO
A highly selective series of inhibitors of the class I phosphatidylinositol 3-kinases (PI3Ks) has been designed and synthesized. Starting from the dual PI3K/mTOR inhibitor 5, a structure-based approach was used to improve potency and selectivity, resulting in the identification of 54 as a potent inhibitor of the class I PI3Ks with excellent selectivity over mTOR, related phosphatidylinositol kinases, and a broad panel of protein kinases. Compound 54 demonstrated a robust PD-PK relationship inhibiting the PI3K/Akt pathway in vivo in a mouse model, and it potently inhibited tumor growth in a U-87 MG xenograft model with an activated PI3K/Akt pathway.
Assuntos
Classe I de Fosfatidilinositol 3-Quinases/antagonistas & inibidores , Piperazinas/síntese química , Piridinas/síntese química , Sulfonamidas/síntese química , Triazinas/síntese química , Animais , Disponibilidade Biológica , Classe I de Fosfatidilinositol 3-Quinases/fisiologia , Cristalografia por Raios X , Desenho de Fármacos , Feminino , Humanos , Indazóis/síntese química , Indazóis/farmacocinética , Indazóis/farmacologia , Camundongos , Camundongos Nus , Microssomos Hepáticos/metabolismo , Modelos Moleculares , Piperazinas/farmacocinética , Piperazinas/farmacologia , Proteínas Proto-Oncogênicas c-akt/fisiologia , Purinas/síntese química , Purinas/farmacocinética , Purinas/farmacologia , Pirazóis/síntese química , Pirazóis/farmacocinética , Pirazóis/farmacologia , Piridinas/farmacocinética , Piridinas/farmacologia , Pirimidinas/síntese química , Pirimidinas/farmacocinética , Pirimidinas/farmacologia , Ratos , Transdução de Sinais , Relação Estrutura-Atividade , Sulfonamidas/farmacocinética , Sulfonamidas/farmacologia , Sulfonas/síntese química , Sulfonas/farmacocinética , Sulfonas/farmacologia , Serina-Treonina Quinases TOR/antagonistas & inibidores , Triazinas/farmacocinética , Triazinas/farmacologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Primary health care in Australia has undergone 2 decades of change. Starting with a vision for a national health strategy with general practice at its core, Australia established local meso-level primary health care organizations--Divisions of General Practice--moving from focus on individual practitioners to a professional collective local voice. The article identifies how these meso-level organizations have helped the Australian primary health care system evolve by supporting the roll-out of initiatives including national practice accreditation, a focus on quality improvement, expansion of multidisciplinary teams into general practice, regional integration, information technology adoption, and improved access to care. Nevertheless, there are still challenges to ensuring equitable access and the supply and distribution of a primary care workforce, addressing the increasing rates of chronic disease and obesity, and overcoming the fragmentation of funding and accountability in the Australian system.
Assuntos
Reforma dos Serviços de Saúde/organização & administração , Programas Nacionais de Saúde/organização & administração , Atenção Primária à Saúde/organização & administração , Austrália , Medicina Geral/organização & administração , Informática Médica , Melhoria de Qualidade/organização & administraçãoRESUMO
OBJECTIVE: We aimed to determine the impact of clinic based retinal photography on access to appropriate screening for diabetic retinopathy (DR). DESIGN, SETTING AND PARTICIPANTS: We opportunistically recruited patients undergoing their annual diabetic cycle of care over a two year period in the urban Indigenous primary health care clinic. Data were collected on retinal outcomes, health variables and referral patterns. MAIN OUTCOME MEASURES: Access to appropriate screening and ophthalmic follow up, prevalence of DR, acceptability and feasibility of clinic-based retinal photography were the main outcome measures of this study. RESULTS: One hundred and thirty-two of a possible 147 patients consented to participate. 30% of participants had DR. Appropriate screening and ophthalmic follow up increased six fold, from 20 to 124 participants, following the introduction of the retinal camera. Most participants felt very positive about DR screening. CONCLUSIONS: Primary care DR screening using retinal photography can improve access to DR screening for indigenous patients, reduce the burden on busy outpatient departments and should reduce visual loss. Policy-makers could contribute to screening sustainability by funding a medicare item-number for primary care based DR screening associated with the annual diabetic cycle of care. An upfront Practice Incentive Program (PIP) payment could offset set up costs.
Assuntos
Retinopatia Diabética/diagnóstico , Programas de Rastreamento/métodos , Havaiano Nativo ou Outro Ilhéu do Pacífico/estatística & dados numéricos , Fotografação , Atenção Primária à Saúde/organização & administração , Adulto , Idoso , Austrália/epidemiologia , Complicações do Diabetes/etnologia , Retinopatia Diabética/complicações , Retinopatia Diabética/etnologia , Feminino , Seguimentos , Acessibilidade aos Serviços de Saúde , Humanos , Masculino , Pessoa de Meia-Idade , Aceitação pelo Paciente de Cuidados de Saúde , Prevalência , Encaminhamento e Consulta/estatística & dados numéricos , Sensibilidade e Especificidade , População Urbana , Adulto JovemRESUMO
BACKGROUND: Type 2 Diabetes Mellitus is one of the most disabling chronic conditions worldwide, resulting in significant human, social and economic costs and placing huge demands on health care systems. The Inala Chronic Disease Management Service aims to improve the efficiency and effectiveness of care for patients with type 2 diabetes who have been referred by their general practitioner to a specialist diabetes outpatient clinic. Care is provided by a multidisciplinary, integrated team consisting of an endocrinologist, diabetes nurse educators, General Practitioner Clinical Fellows (general practitioners who have undertaken focussed post-graduate training in complex diabetes care), and allied health personnel (a dietitian, podiatrist and psychologist). METHODS/DESIGN: Using a geographical control, this evaluation study tests the impact of this model of diabetes care provided by the service on patient outcomes compared to usual care provided at the specialist diabetes outpatient clinic. Data collection at baseline, 6 and 12-months will compare the primary outcome (glycaemic control) and secondary outcomes (serum lipid profile, blood pressure, physical activity, smoking status, quality of life, diabetes self-efficacy and cost-effectiveness). DISCUSSION: This model of diabetes care combines the patient focus and holistic care valued by the primary care sector with the specialised knowledge and skills of hospital diabetes care. Our study will provide empirical evidence about the clinical effectiveness of this model of care. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry ACTRN12608000010392.
Assuntos
Diabetes Mellitus Tipo 2/terapia , Serviços Preventivos de Saúde/métodos , Idoso , Doença Crônica , Protocolos Clínicos , Assistência Integral à Saúde/organização & administração , Coleta de Dados , Prestação Integrada de Cuidados de Saúde , Complicações do Diabetes/diagnóstico , Diabetes Mellitus Tipo 2/complicações , Gerenciamento Clínico , Feminino , Pesquisa sobre Serviços de Saúde , Humanos , Masculino , Pessoa de Meia-Idade , Avaliação de Resultados em Cuidados de Saúde , Avaliação de Programas e Projetos de Saúde , Queensland , Autocuidado , Fumar/epidemiologiaRESUMO
BACKGROUND & AIMS: Infliximab induces immune cell apoptosis by outside-to-inside signaling through transmembrane tumor necrosis factor-alpha (mTNF). However, in inflamed gut, myofibroblasts also produce TNF-alpha, and the affects of anti-TNF antibodies on these structural cells are unknown. We investigated the action of infliximab on apoptosis, the production of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMP)-1, and migration of Crohn's disease (CD) myofibroblasts. METHODS: Colonic myofibroblasts were isolated from patients with active CD and controls. mTNF was evaluated by Western blotting and flow cytometry. Infliximab-treated myofibroblasts were analyzed for apoptosis by Annexin V staining and caspase-3. TIMP-1 and MMPs were measured by Western blotting, and fibroblast migration was assessed by using an in vitro wound-healing scratch assay. RESULTS: CD myofibroblasts showed higher mTNF expression than control myofibroblasts. Infliximab had no effect on CD myofibroblast apoptosis, caspase-3 activation, and production of MMP-3 and MMP-12. However, infliximab induced a significant dose-dependent increase in TIMP-1 production, which was inhibited by the p38 mitogen-activated protein kinase inhibitor SB 203580. The anti-TNF agents adalimumab, etanercept, and p55 TNF-receptor-human IgG fusion protein also increased TIMP-1 production. The migration of CD myofibroblasts was enhanced significantly by infliximab and recombinant human TIMP-1, and infliximab-induced migration was inhibited by anti-TIMP-1 neutralizing antibody. Infliximab also decreased CD myofibroblast collagen production. CONCLUSIONS: Our findings show a novel therapeutic pathway for anti-TNF therapies in enhancing TIMP-1 production and myofibroblast migration, which may reduce MMP activity and facilitate the wound healing.
Assuntos
Autoanticorpos/farmacologia , Doença de Crohn/imunologia , Doença de Crohn/patologia , Fibroblastos/imunologia , Fator de Necrose Tumoral alfa/imunologia , Adulto , Anti-Inflamatórios/farmacologia , Anticorpos Monoclonais/farmacologia , Apoptose/efeitos dos fármacos , Apoptose/imunologia , Autoanticorpos/imunologia , Biópsia , Células Cultivadas , Colágeno/metabolismo , Colo/imunologia , Colo/patologia , Fibroblastos/metabolismo , Fibroblastos/patologia , Humanos , Infliximab , Metaloproteinase 12 da Matriz/metabolismo , Metaloproteinase 3 da Matriz/metabolismo , Pessoa de Meia-Idade , Técnicas de Cultura de Órgãos , Inibidor Tecidual de Metaloproteinase-1/imunologia , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Inibidor Tecidual de Metaloproteinase-1/farmacologia , Cicatrização/efeitos dos fármacos , Cicatrização/imunologiaRESUMO
This study examined the suitability of microdialysis to assess the time course of cytokine generation from discrete sites within the skin following intradermal injection of allergen. Cytokines were recovered using two microdialysis probes, one close to the point of allergen injection and the other 1 cm away but within the area of the late-phase induration. Skin biopsies taken at both sites were stained immunocytochemically to investigate possible relationships between cytokine generation, expression of adhesion molecules, and recruitment of neutrophils and eosinophils during the late-phase allergic response. The cytokine response to probe insertion was assessed using a single probe in the opposite arm (control). At baseline, microdialysate contained low levels of IL-1alpha, IL-5, IL-8, IL-12, GM-CSF, and TNFalpha (n=27-33). At control sites, this was followed by increases in IL-6 and IL-8 at 3 and 6 hours. Allergen increased TNFalpha levels in 3/11 individuals within 30 minutes at the injection site. Levels of IL-6 and IL-8 rose rapidly and were significantly greater (P<0.05) than that of controls at 3 and 6 hours at both injection and distant sites. Adhesion molecule expression and leukocyte infiltration were elevated only at the allergen injection site, suggesting a complex relationship between cytokine generation and cellular events in allergic inflammation. In conclusion, microdialysis can be used to distinguish temporal and spatial changes in protein profiles in the skin. Furthermore, when used in conjunction with skin biopsies, it provides novel information about the mechanisms of dermal inflammation.
Assuntos
Alérgenos/metabolismo , Citocinas/metabolismo , Microdiálise/métodos , Pele/metabolismo , Adesão Celular , Edema/patologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Humanos , Inflamação , Interleucina-12/metabolismo , Interleucina-1alfa/metabolismo , Interleucina-5/metabolismo , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Pele/patologia , Fenômenos Fisiológicos da Pele , Fatores de Tempo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
The Brisbane South Centre for Health Services Integration (BSCHSI) initiative used a collocation strategy to integrate local service delivery across three different health organisations. Physical collocation was combined with validated integration strategies to improve organisational operations among five different work teams involving 90 different individuals. Enhanced communication, increased knowledge of collocating groups, and the development of collaboration and partnerships were key positive outcomes.
Assuntos
Planejamento em Saúde Comunitária/organização & administração , Comportamento Cooperativo , Atitude do Pessoal de Saúde , Grupos Focais , Humanos , Programas Nacionais de Saúde , Queensland , Inquéritos e QuestionáriosRESUMO
The Brisbane Inner South E-referral Project (BISEP) developed an application which allowed general practitioners, from their desktop, to successfully search for and book an available hospital outpatient appointment for patients with suspected cancer, send the referral electronically, and inform the patient of both the appointment and referral during the consultation. The hospital changed their outpatient department processes to allow such functionality for local GPs with patients with suspected cancer, working from a mutually agreed set of best practice referral criteria. A group of 19 GPs participated in an 11-week pilot implementation of the application, and were enthusiastic about continuing and expanding the approach. Patient satisfaction measures post intervention indicated that they perceived no major disadvantage in this form of outpatient department referral.
Assuntos
Agendamento de Consultas , Medicina de Família e Comunidade/organização & administração , Internet , Ambulatório Hospitalar/organização & administração , Encaminhamento e Consulta/organização & administração , Integração de Sistemas , Humanos , Projetos Piloto , QueenslandRESUMO
Studies in mice indicate that alpha-melanocyte-stimulating hormone (alphaMSH) is immunosuppressive, but it is not known whether alphaMSH suppresses human immune responses to exogenous Ags. Human PBMCs, including monocytes, express the melanocortin 1 receptor (MC1R), and it is thought that the ability of alphaMSH to alter monocyte-costimulatory molecule expression and IL-10 release is mediated by this receptor. However, the MC1R gene is polymorphic, and certain MC1R variants compromise receptor signaling via cAMP, resulting in red hair and fair skin. Here, we have investigated whether alphaMSH can suppress Ag-induced lymphocyte proliferation in humans and whether these effects are dependent on MC1R genotype. alphaMSH suppressed streptokinase-streptodornase-induced lymphocyte proliferation, with maximal inhibition at 10(-13)-10(-11) M alphaMSH. Anti-IL-10 Abs failed to prevent suppression by alphaMSH, indicating that it was not due to MC1R-mediated IL-10 release by monocytes. Despite variability in the degree of suppression between subjects, similar degrees of alphaMSH-induced immunosuppression were seen in individuals with wild-type, heterozygous variant, and homozygous/compound heterozygous variant MC1R alleles. RT-PCR of streptokinase-streptodornase-stimulated PBMCs for all five melanocortin receptors demonstrated MC1R expression by monocytes/macrophages, MC1R and MC3R expression by B lymphocytes, but no melanocortin receptor expression by T lymphocytes. In addition, alphaMSH did not significantly inhibit anti-CD3 Ab-induced lymphocyte proliferation, whereas alphaMSH and related analogs (SHU9119 and MTII) inhibited Ag-induced lymphocyte proliferation in monocyte-depleted and B lymphocyte-depleted assays. These findings demonstrate that alphaMSH, acting probably via MC1R on monocytes and B lymphocytes, and possibly also via MC3R on B lymphocytes, has immunosuppressive effects in humans but that suppression of Ag-induced lymphocyte proliferation by alphaMSH is independent of MC1R gene status.